Novel HSP90 Inhibitor

ABSTRACT

Disclosed is a triazole derivative(s) represented by the general formula (1) below or a pharmacologically acceptable salt(s) thereof. Also disclosed are a prodrug(s) of such a triazole derivative(s) and an HSP90 inhibitor(s) containing any one of them as an active constituent. (1) (In the formula, X represents a halogen atom, an optionally substituted alkyl group, an optionally substituted alkynyl group or the like; Y represents a mercapto group, a hydroxyl group, an optionally substituted sulfonyl group, an optionally substituted amino group or the like; and R represents an optionally substituted aryl or alkyl group or the like.)

This application is a continuation of U.S. Ser. No. 11/885,575, filedSep. 4, 2007, which is a §371 of PCT/JP2006/304496 filed Mar. 8, 2006,the disclosures of which are hereby incorporated by reference.

TECHNICAL FIELD

The present invention relates to a novel triazole derivative and anHSP90 inhibitor containing the triazole derivative as an activeingredient. The triazole derivative of the present invention inhibitsthe function of HSP90 by binding to its ATP binding site, blocks thebinding of HSP90 and its client protein and finally suppresses cellulargrowth.

BACKGROUND ART

Molecular chaperones are a general term for proteins that form a complextemporally with client proteins to promote the formation of theconformation of the client proteins. These proteins, the activity ofwhich is to help folding and association of protein and to preventaggregation, are broadly defined as molecular chaperones and classifiedinto several families according to their molecular weights (HSP90,HSP70, HSP60, HSP40, small HSPs and the like). In particular, HSP90 hasbeen known to interact with many molecules which are involved in theintracellular signal transduction, and it is becoming clear that HSP90is deeply involved in cell cycle regulation, and carcinogenesis, growthand survival signal of cells.

HSP90 is a molecular chaperone present in cells in abundance (occupies1-2% of total soluble protein), distributed in the cytoplasm evenly andexists mainly as dimers. The activity of HSP90 alone in protein foldingis low, and HSP90 functions cooperatively with other molecularchaperones having a folding activity (hereinafter called co-chaperones)such as HSP70 and p23. HSP90 is often needed for its function of clientproteins that form a complex, and the action mechanism is based on thebiochemical characteristic that HSP90 specifically recognizes a proteinunder the condition of unstable folding and binds to it. HSP90 performsATP dependent folding (re-folding) of a denatured protein or a proteinthat is not folded. Especially, it is needed for constructing thestructure of various key proteins (steroid receptors, Raf serinekinases, tyrosine kinases) which are involved in cancer related signaltransduction. According to the recent findings, the control function ofmany key signal molecules is lost in human tumors, and these requireHSP90 to maintain the function (non-patent document 1).

Geldanamycin (hereinafter called GM) is an ansamycin natural product,which was initially discovered in microorganisms as a tyrosine kinaseinhibitor, but its direct inhibitory effect on a tyrosine kinase waslow, and later it was found that this drug acted on HSP90 specifically.Radicicol (hereinafter called RD) is a macrolide natural product which,as a different structure from GM, also acts on HSP90 and inhibits itsfunction. It has been known that GM and RD induce degradation of variouskey proteins (steroid receptor, Raf, Her2 and the like) which areinvolved in signal transduction related to cancer and cause growthinhibition of various cancer cells in vitro. HSP90 contains at theN-terminal an ATP/ADP binding site which plays an important role incontrolling the chaperone function. This site is specific for and wellpreserved in the HSP90 family, and does not exist in other molecularchaperones. It has been elucidated by crystallographic analysis that GMand RD directly bind to this ATP/ADP binding site as antagonists(non-patent documents 2 and 3). It is also known that these antagonistsinhibit the association with a co-chaperone such as p23 by binding tothe ATP/ADP binding site. As the result, the composition of thechaperone complex which contains client proteins and HSP90 is changed,and eventually the client proteins are released from the complex anddegraded mainly in the ubiquitin-proteasome pathway. Thus, theantiproliferative action on cancer cells by HSP90 antagonists will becaused by a depression of the client protein of HSP90 and the blockingof signal transduction pathway by the depression.

The HSP90 antagonist acts selectively on client proteins folded intoHSP90, and does not affect the function and the amount of expression ofother proteins at all. Studies have shown that in the process ofcarcinogenesis, a plurality of gene abnormalities are accumulated, andin many tumor cells, mutated proteins require more of the chaperoneactivity than normal proteins. HSP90 is overexpressed in variouscancers. From the analyses of pharmacokinetics of a GM derivative,17-AAG, in animal models, more of the 17-AAG is accumulated in cancer incomparison to the normal cells. From these reports, it is expected thatthe HSP90 antagonist acts on cancer cells specifically, not on normalcells. Also, since cancer cells under a kind of stressful condition suchas abnormal protein expression, low oxygen and nutritional starvationare dependent on HSP90 at a higher degree, it would appear that thesensitivity of cancer cells against the HSP90 antagonist is higher.

Among the HSP90 antagonists, 17-AAG is subjected to ongoing Phase I/IIclinical trials, and investigations on RD derivatives are also beingconducted (non-patent document 4), but any one of these has problems foruse as a pharmaceutical product in physical properties such as molecularweight, stability, and water solubility. A water soluble and lowmolecular weight HSP90 inhibitor is sought as a useful pharmaceuticalproduct. An adenine derivative, PU3 and its derivatives have beenreported to be a low molecular weight HSP90 inhibitor (Patent Document1, non-patent document 5, non-patent document 6 and non-patent document7). Also, a 1,3-dihydroxybenzene derivative, to which a 5-member ring isbound, has been reported to be a HSP90 inhibitor (patent document 2,patent document 3, patent document 4, patent document 5, patent document6, and patent document 7), but the antiproliferative activity againstcancer cells in vitro is weak (patent document 2). Further, the patentdocument 8 describes benzene derivatives, to which a 5-member ring isbound, as an antagonist of HSP90, but data of the HSP90 inhibitoryactivity of a derivative in which the 5-member ring has a triazoleskeleton is not disclosed. On the other hand, the fact that the triazolederivatives of the present invention have HSP90 inhibitory activity isnot known in literature.

-   Patent Document 1: International Publication No. 02/036075-   Patent Document 2: International Publication No. 03/055860-   Patent Document 3: International Publication No. 04/050087-   Patent Document 4: International Publication No. 04/056782-   Patent Document 5: International Publication No. 04/096212-   Patent Document 6: International Publication No. 04/072051-   Patent Document 7: International Publication No. 05/000300-   Patent Document 8: International Publication No. 05/041879-   Non-Patent Document 1: Hsp90 inhibitors as novel cancer    chemotherapeutic agents. Trends Mol. Med. 2002; 8(4 Suppl.): p.    S55-61.-   Non-Patent Document 2: Inhibition of heat shock protein HSP90-pp    60v-src heteroprotein complex formation by benzoquinone ansamycins:    essential role for stress proteins in oncogenic transformation. Proc    Natl Acad Sci U.S.A. 1994; 91(18): p8324-8328.-   Non-Patent Document 3: Crystal structure of an Hsp90-geldanamycin    complex: targeting of a protein chaperone by an antitumor agent.    Cell 1997 Apr. 18; 89(2): p. 239-250.-   Non-Patent Document 4: The clinical applications of heat shock    protein inhibitors in cancer present and future. Curr. Cancer Drug    Targets. 2003 October; 3(5): p. 385-390.-   Non-Patent Document 5: A small molecule designed to bind to the    adenine nucleotide pocket of Hsp90 causes Her2 degradation and the    growth arrest and differentiation of breast cancer cells. G. Chiosis    et al., Chem. Biol. 2001 March; 8(3): p. 289-299.-   Non-Patent Document 6: Targeting Wide-Range Oncogenic Transformation    via PU24FC1, a specific Inhibitor of Tumor Hsp90. M. Vilenchik et    al., Chem. Bio., 11, p. 787-797 (2004).-   Non-Patent Document 7: Adenine derived inhibitors of the molecular    chaperone HSP90-SAR explained through multiple X-ray structures. D.    Dymock et al., Bioorg. Med. Chem. Lett., 14 (02), p. 325-328 (2004).

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

The inhibitors of HSP90, which is involved in cell growth, are expectedto be effective against cancer cells selectively as described above, andsome of them have been under development, but so far no inhibitor havingsufficient stability and efficacy required for pharmaceutical drugs hasbeen obtained, and a HSP90 inhibitor usable as a drug is desired.

To solve the problem described above, the present inventors investigatedrigorously and as the result discovered that a triazole derivativerepresented by the following general formula (1),

a prodrug thereof or a pharmaceutically acceptable salt thereof inhibitsHSP90 to complete the present invention.

That is, the present invention relates to

(1) a triazole derivative represented by the following general formula(1) or a pharmaceutically acceptable salt thereof,

wherein N represents a nitrogen atom; X represents a mercapto group,hydroxy group, halogen atom, nitro group, cyano group, an optionallysubstituted alkyl group, an optionally substituted alkenyl group, anoptionally substituted alkynyl group, optionally substituted carbocyclicor heterocyclic aryl group, an optionally substituted alkylthio group,an optionally substituted arylthio group, an optionally substitutedalkylsulfinyl group, an optionally substituted arylsulfinyl group, anoptionally substituted alkylsulfonyl group, an optionally substitutedarylsulfonyl group, an optionally substituted sulfamoyl group, anoptionally substituted alkoxyl group, an optionally substituted aryloxygroup, an optionally substituted acyloxy group, an optionallysubstituted alkoxycarbonyloxy group, an optionally substitutedcarbamoyloxy group, an optionally substituted amino group, an optionallysubstituted acylamino group, an optionally substitutedalkoxycarbonylamino group, an optionally substituted ureido group, anoptionally substituted sulfonylamino group, an optionally substitutedsulfamoylamino group, a formyl group, an optionally substituted acylgroup, an optionally substituted carboxyl group, alkoxycarbonyl group,an optionally substituted carbamoyl group, an optionally substituted oran optionally substituted silyl group; Y represents a mercapto group,hydroxy group, halogen atom, cyano group, sulfonyl group, an optionallysubstituted alkylthio group, an optionally substituted arylthio group,an optionally substituted alkylsulfinyl group, an optionally substitutedarylsulfinyl group, an optionally substituted sulfamoyl group, anoptionally substituted alkoxyl group, an optionally substituted aryloxygroup, an optionally substituted acyloxy group, an optionallysubstituted alkoxycarbonyloxy group, an optionally substitutedcarbamoyloxy group, an optionally substituted amino group, an optionallysubstituted acylamino group, an optionally substitutedalkoxycarbonylamino group, an optionally substituted ureido group, anoptionally substituted sulfonylamino group, an optionally substitutedsulfamoylamino group, a formyl group, an optionally substituted acylgroup or an optionally substituted silyl group; R represents anoptionally substituted carbocyclic or heterocyclic aryl group, or anoptionally substituted alkyl group, an optionally substituted alkenylgroup, an optionally substituted alkynyl group or an optionallysubstituted amino group;(2) The triazole derivative according to the aforementioned (1), whereinX is located at the 5 position of a 2,4-dihydroxyphenyl group whichbinds to triazole ring at 1 position in the general formula (1) of theaforementioned (1), or a pharmaceutically acceptable salt thereof;(3) The triazole derivative according to the aforementioned (1) or (2),wherein X is an optionally substituted alkyl group, an optionallysubstituted alkenyl group or an optionally substituted alkynyl group, ora halogen atom in the general formula I of the aforementioned (1), or apharmaceutically acceptable salt thereof;(4) The triazole derivative according to any one of the aforementioned(1) to (3), wherein the compound represented by the general formula (1)of the aforementioned (1) is an acetylene derivative represented by thefollowing general formula (1-1) or a pharmaceutically acceptable saltthereof;

wherein R and Y represent the same meanings as in R and Y of the generalformula (1) of the aforementioned (1), X^(a) represents an optionallysubstituted methylene group, n represents an integer from 0 to 3, X^(b)represents a hydrogen atom, an optionally substituted alkyl group, anoptionally substituted alkenyl group or an optionally substitutedalkynyl group, an optionally substituted carbocyclic or heterocyclicaryl group, halogen atom, sulfamoyl group, formyl group, acyl group,carboxyl group, carbamoyl group or silyl group;(5) The triazole derivative according to the aforementioned (4) or apharmaceutically acceptable salt thereof, wherein n is 1 in the generalformula (1-1) of the aforementioned (4);(6) The triazole derivative according to any one of the aforementioned(1) to (5) or a pharmaceutically acceptable salt thereof, wherein Y isany of a mercapto group, hydroxy group, an optionally substitutedsulfonyl group or alkylthio group in the general formula (1) of theaforementioned (1) or the general formula (1-1) of the aforementioned(4);(7) The triazole derivative according to any one of the aforementioned(1) to (6) or a pharmaceutically acceptable salt thereof, wherein Y isan alkylsulfonyl group optionally substituted on the alkyl groupthereof, or an arylsulfonyl group optionally substituted on the arylgroup thereof in the general formula (1) of the aforementioned (1) orthe general formula (1-1) of the aforementioned (4);(8) The triazole derivative according to any one of the aforementioned(1) to (6) or a pharmaceutically acceptable salt thereof, wherein Y is amercapto group in the general formula (1) of the aforementioned (1) orthe general formula (1-1) of the aforementioned (4);(9) The triazole derivative according to any one of the aforementioned(1) to (6) or a pharmaceutically acceptable salt thereof, wherein Y is ahydroxy group in the general formula (1) of the aforementioned (1) orthe general formula (1-1) of the aforementioned (4);(10) The triazole derivative according to any one of the aforementioned(1) to (9) or a pharmaceutically acceptable salt thereof, wherein R isan optionally substituted carbocyclic or heterocyclic aryl group in thegeneral formula (1) of the aforementioned (1) or the general formula(1-1) of the aforementioned (4);(11) The triazole derivative according to any one of the aforementioned(1) to (10) or a pharmaceutically acceptable salt thereof, wherein R isrepresented by the following general formula (2) in the general formula(1) of the aforementioned (1),

wherein m is an integer from 0 to 5, A is an optionally substitutedcyclic or non-cyclic amino group, an optionally substituted cyclic ornon-cyclic acylamino group or an optionally substituted cyclic ornon-cyclic sulfonylamino group;(12) The triazole derivative according to the aforementioned (11) or apharmaceutically acceptable salt thereof, wherein m is 0 or 1 and A is acyclic amino group in the general formula (2) of the aforementioned(11);(13) The triazole derivative according to any one of the aforementioned(1) to (10) or a pharmaceutically acceptable salt thereof, wherein R isrepresented by the following general formula (2-2) in the generalformula (1) of the aforementioned (1) or the general formula (1-1) ofthe aforementioned (4),

wherein n^(a) is an integer from 1 to 5, A^(a) is an optionallysubstituted alkyl group having carbons from 1 to 6 and when n^(a) is 2to 5, the neighboring substituents taken together may form a ring;(14) The triazole derivative according to any one of the aforementioned(1) to (9) or a pharmaceutically acceptable salt thereof, wherein R isan optionally substituted alkyl group in the general formula (1) of theaforementioned (1) or the general formula (1-1) of the aforementioned(4);(15) The triazole derivative according to any one of the aforementioned(1) to (12) or a pharmaceutically acceptable salt thereof, wherein thecompound represented by the general formula (1) of the aforementioned(1) is represented by the following general formula (4),

wherein X represents a chlorine atom, ethyl group, isopropyl group,tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group or2-butynyl group; Y represents a mercapto group, an optionallysubstituted alkylsulfonyl group or hydroxy group; m is 0 or 1; and Arepresents a cyclic amino group;(16) The triazole derivative according to any one of the aforementioned(1) to (10) and (15), or a pharmaceutically acceptable salt thereof,wherein the compound represented by the general formula (1) of theaforementioned (1) is represented by the following general formula(1-2),

wherein X represents a chlorine atom, ethyl group, isopropyl group,tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group or2-butynyl group; Ar^(a) represents a 4-methoxyphenyl group,3-methoxyphenyl group, 3,4-dimethoxyphenyl group, 3,4,5-trimethoxyphenylgroup or 3,4-methylenedioxyphenyl group;(17) The triazole derivative according to any one of the aforementioned(1) to (10) and (14), or a pharmaceutically acceptable salt thereof,wherein the compound represented by the general formula (1) of theaforementioned (1) is represented by the following general formula(1-3),

wherein X represents a chlorine atom, ethyl group, isopropyl group,tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group or2-butynyl group; Alk represents an optionally substituted alkyl group;(18) The triazole derivative according to the aforementioned (1) or apharmaceutically acceptable salt thereof, which is selected from thegroup consisting of:

-   4-isopropyl-6-{5-mercapto-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    (SH-a01),-   4-isopropyl-6-{5-mercapto-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    (SH-a02),-   4-[4-(4-bromo-phenyl)-5-mercapto-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol    (SH-a03),-   4-{5-hydroxy-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol    (OH-a01),-   4-{5-hydroxy-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol    (OH-a02),-   5-[5-(but-2-ynyl)-2,4-dihydroxy-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-c02),-   4-(but-2-ynyl-2-yl)-6-{5-mercapto-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    (SH-c02),-   4-bromo-6-{5-mercapto-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    (SH-d01),-   4-isopropyl-6-{5-methanesulfonyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    (SFN-a02),-   4-isopropyl-6-[5-methylsulfinyl-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol    (SFX-a08),-   4-isopropyl-6-[5-methanesulfonyl-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol    (SFN-a08),-   5-[2,4-dihydroxy-5-(prop-2-ynyl)-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-e02),-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a08),-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(3-methoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a09),-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(3,4-dimethoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a10),-   4-[benzo[1,3]dioxol-5-yl]-5-(2,4-dihydroxy-5-isopropyl-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one,-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-hydroxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a11),-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[2-(morpholin-4-yl)-pyrimidin-5-yl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a17),-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a13),-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-isopropyl-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a21),-   4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol    (SFN2-a08),-   4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(3-piperidin-1-yl-propane-1-sulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol    (SFN3-a08), and-   N-[5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-methane    sulfonamide (N1-a08).    (19) A prodrug of the triazole derivative according to any one of    the aforementioned (1) to (18) or a pharmaceutically acceptable salt    thereof;    (20) A medicine containing as an active ingredient the prodrug of    the triazole derivative according to any one of the    aforementioned (1) to (18) or a pharmaceutically acceptable salt of    the prodrug;    (21) An HSP90 inhibitor containing as an active ingredient the    triazole derivative according to any one of the aforementioned (1)    to (18), a prodrug thereof or a pharmaceutically acceptable salt    thereof; and    (22) An anticancer agent containing as an active ingredient the    triazole derivative according to any one of the aforementioned (1)    to (18), a prodrug thereof or a pharmaceutically acceptable salt    thereof.

Effect of the Invention

The present invention can provide a drug composition containing acompound having a superior HSP90 inhibitory activity or apharmaceutically acceptable salt thereof as an active ingredient, inparticular a therapeutic agent for cancer.

BEST MODE FOR CARRYING OUT THE INVENTION

Following is the detailed description of the present invention.

In the present invention, a halogen atom indicates a fluorine atom,chlorine atom, bromine atom or iodine atom.

In the present invention, an alkyl group means, unless specifiedotherwise, a linear, branched or cyclic alkyl group having 1-20 carbons,preferably 1-8 carbons. The linear alkyl group includes, for example, amethyl group, ethyl group, propyl group, n-butyl group, n-pentyl group,and n-hexyl group. The branched alkyl group includes, for example, anisopropyl group, tert-butyl group, and 2,2-dimethylpropyl group. Thecyclic alkyl group includes, for example, a cyclopropyl group,cyclobutyl group, cyclopentyl group, cyclohexyl group, and adamantylgroup.

In the present invention, an alkenyl group means a linear, branched orcyclic alkenyl group having a carbon-carbon double bond at one or moreplaces and having 2-20 carbons, preferably 2-8 carbons. The linearalkenyl group includes, for example: a 1-alkenyl group such as anethenyl group, 1-propenyl group, and 1-butenyl group; and a 2-alkenylgroup such as a 2-butenyl group, and 2-pentenyl group. The branchedalkenyl group includes, for example, an isopropenyl group,3-methyl-1-butenyl group, and geranyl group.

In the present invention, an alkynyl group means an alkynyl group havinga carbon-carbon triple bond at one or more places and having 2-20carbons, preferably 2-8 carbons. Examples include: a 1-alkynyl groupsuch as ethynyl group, 1-propynyl group, and 3,3-dimethyl-1-butynylgroup; and a 2-alkynyl group such as a 2-propynyl group, 2-butynylgroup, 3-phenyl-2-propynyl group, 4,4-dimethyl-2-pentynyl group, and3-trimethylsilyl-2-propynyl group.

In the present invention, a carbocyclic aryl group includes, forexample, a phenyl group, and naphthyl group. A heterocyclic aryl groupincludes, for example, a pyridyl group, pyrimidinyl group, quinolylgroup, quinazolyl group, naphthyridinyl group, furyl group, pyrrolylgroup, indolyl group, imidazolyl group, pyrazolyl group, oxazolyl group,isoxazolyl group, and triazolyl group.

In the present invention, when the description “an optionallysubstituted” is used, the substituent includes, for example, a hydrogenatom, mercapto group, hydroxy group, halogen atom, nitro group, cyanogroup, alkyl group, alkenyl group, alkynyl group, carbocyclic orheterocyclic aryl group, alkylthio group, arylthio group, alkylsulfinylgroup, arylsulfinyl group, alkylsulfonyl group, arylsulfonyl group,sulfamoyl group, alkoxyl group, aryloxy group, acyloxy group,alkoxycarbonyloxy group, carbamoyloxy group, substituted ornon-substituted amino group, acylamino group, alkoxycarbonylamino group,ureido group, sulfonylamino group, sulfamoylamino group, formyl group,acyl group, carboxyl group, alkoxycarbonyl group, carbamoyl group, andsilyl group. The position of substitution on an aromatic ring may beortho, meta or para.

In the present invention, an alkylthio group means the alkylthio grouphaving 1-8 carbons and includes, for example, a methylthio group,isopropylthio group, and benzylthio group. An arylthio group includes,for example, a phenylthio group, naphthylthio group, and pyridylthiogroup. An alkylsulfinyl group means the alkylsulfinyl group having 1-8carbons and includes, for example, a methylsulfinyl group,isopropylsulfinyl group, and benzylsulfinyl group. An arylsulfinyl groupincludes, for example, phenylsulfinyl group, naphthylsulfinyl group, andpyridylsulfinyl group. An optionally substituted sulfonyl groupincludes, for example, an alkylsulfonyl group, alkenylsulfonyl group,alkynylsulfonyl group, and arylsulfonyl group. An alkylsulfonyl groupmeans the alkylsulfonyl group having 1-8 carbons and includes, forexample, a methylsulfonyl group, isopropylsulfonyl group, andbenzylsulfonyl group. An arylsulfonyl group includes, for example, aphenylsulfonyl group, naphthylsulfonyl group, and pyridylsulfonyl group.A sulfamoyl group includes, for example, a dimethylsulfamoyl group, andphenylsulfamoyl group.

In the present invention, an alkoxyl group means the alkoxyl grouphaving 1-8 carbons and includes, for example, a methoxyl group,isopropoxyl group, and benzyloxy group. An aryloxy group includes, forexample, a phenoxyl group, naphthyloxy group, and pyridyloxy group. Anacyloxy group means the acyloxy group having 1-8 carbons and includes,for example, an acetoxyl group, and benzoyloxy group. Analkoxycarbonyloxy group means the alkoxycarbonyloxy group having 1-8carbons and includes, for example, a methoxycarbonyloxy group, andtrifluoromethoxycarbonyl group. A carbamoyloxy group includes, forexample, a dimethylcarbamoyloxy group, and phenylcarbamoyloxy group.

In the present invention, an amino group includes, for example, anon-substituted amino group, dimethylamino group, morpholino group,piperidinyl group, 4-methylpiperazine-1-yl group, and phenylamino group.An acylamino group includes, for example, an acetylamino group, andbenzoylamino group. An alkoxycarbonylamino group includes, for example,a methoxycarbonylamino group, ethoxycarbonylamino group, andbenzyloxycarbonylamino group. An ureido group includes, for example, atrimethylureido group, and 1-methyl-3-phenyl-ureido group. Asulfonylamino group includes, for example, a methanesulfonylamino group,and benzenesulfonylamino group. A sulfamoylamino group includes, forexample, a dimethylsulfamoylamino group.

In the present invention, an acyl group includes, for example, an acetylgroup, pivaloyl group, benzoyl group, and pyridinecarbonyl group. Analkoxycarbonyl group includes, for example, a methoxycarbonyl group, andbenzyloxycarbonyl group. A carbamoyl group includes for example, adimethylcarbamoyl group, and phenylcarbamoyl group.

In the present invention, a silyl group includes, for example, atrimethylsilyl group, triisopropylsilyl group, andtert-butyl-diphenyl-silyl group.

In the present invention, R represents an optionally substitutedcarbocyclic or heterocyclic aryl group or an optionally substitutedalkyl group, an optionally substituted alkenyl group or an optionallysubstituted alkynyl group or substituted or non-substituted amino group.An optionally substituted carbocyclic aryl group represented by Rincludes, for example: a phenyl group; bromophenyl group; aminophenylgroup; methylphenyl group; a group represented by the following generalformula (2),

wherein m represents any one of integer 0-5, A represents an optionallysubstituted cyclic or non-cyclic amino group, an optionally substitutedcyclic or non-cyclic acylamino group or an optionally substituted cyclicor non-cyclic sulfonylamino group; and a group represented by thefollowing general formula (2-2),

wherein n^(a) represents any one of integer 1-5, A^(a) represents analkyl group having 1-6 carbons that may have a substituent and, whenn^(a) is 2-5, the neighboring substituents taken together may form aring.

In the substituent represented by the general formula (2), m isespecially preferable to be 0 or 1. A includes an amino group, acylaminogroup, and sulfonylamino group. The amino group includes a cyclic aminogroup, non-cyclic amino group or aromatic amino group. The cyclic aminogroup includes, for example, a morpholino group, piperidinyl group,piperazinyl group, 4-methylpiperazin-1-yl group, and pyrrolidinyl group.The non-cyclic amino group includes, for example, dimethylamino group,isopropylamino group, cyclohexylamino group, 2-hydroxyethylamino group,and 2-methoxyethylamino group. The aromatic amino group includes, forexample, a phenylamino group. The acylamino group includes, for example,an acetylamino group, and benzoylamino group. The sulfonylamino groupincludes, for example, a methanesulfonylamino group, andbenzenesulfonylamino group. Among them, the cyclic amino groups such asthe morpholino group, piperidinyl group, piperazinyl group,4-methylpiperazin-1-yl group, and pyrrolidinyl group are especiallypreferred.

As a substituent represented by the general formula (2),4-(morpholin-4-yl)-phenyl group, 4-(morpholin-4-ylmethyl)-phenyl groupand 4-(4-methyl-piperazin-1-ylmethyl)-phenyl group are especiallypreferred.

Among the substituents represented by the general formula (2-2), n^(a)is preferably 1 and 2. A^(a) includes a methyl group, ethyl group, andmethylene group, and among them, the methyl group is especiallypreferable. As an alkoxyphenyl group represented by the general formula(2-2), a 4-methoxyphenyl group, 3-methoxyphenyl group,3,4-dimethoxyphenyl group and 3,4-methylenedioxyphenyl group arepreferable and especially the 4-methoxyphenyl group is preferable.

A heterocyclic aryl group represented by R includes, for example, apyridyl group, pyrimidinyl group, quinolyl group, quinazolyl group,naphthylidinyl group, furyl group, pyrrolyl group, indolyl group,imidazolyl group, pyrazolyl group, oxazolyl group, isoxazolyl group, andtriazolyl group, and among them, the pyridyl group and pyrimidinyl groupare preferable. A heterocyclic aryl group that has a substituentincludes, for example, 2-morpholin-4-yl-pyrimidin-5-yl group.

An alkyl group represented by R includes, for example: a linear alkylgroup such as a methyl group, ethyl group, propyl group, and butylgroup; a branched alkyl group such as an isopropyl group, 2-methylpropyl group, and tert-butyl group; and a cyclic alkyl group such as acyclopropyl group, cyclobutyl group, cyclopentyl group, and cyclohexylgroup. The alkyl group, represented by R, may have a substituent(s), andthe substituent includes, for example: a hydroxy group; a linear alkoxylgroup such as a methoxy group, and ethoxy group; a cyclic alkoxyl groupsuch as a tetrahydrofuryl group; an amino group such as a morpholinogroup; and a cyclic acylamino group such as a 2-oxo-pyrrolidin-1-ylgroup. Among the alkyl group represented by R, the isopropyl group isespecially preferable.

An amino group represented by R includes, for example: a linear aminogroup such as a dimethylamino group; and a cyclic amino group such as apiperidino group, and morpholino group, and among them, the cyclic aminogroups such as the piperidino group are preferable.

In the present invention, as a substituent represented by R, anoptionally substituted carbocyclic or heterocyclic aryl group and anoptionally substituted alkyl group are preferable. Among them, anoptionally substituted carbocyclic aryl group and represented by thegeneral formulas (2) and (2-2), and an optionally substituted alkylgroup are preferable, and especially a 4-methoxyphenyl group,4-(morpholin-4-yl)-phenyl group, 4-(morpholin-4-ylmethyl)-phenyl group,4-(4-methyl-piperazin-1-ylmethyl)-phenyl group and isopropyl group arepreferable.

In the present invention, a substituent represented by Y includes amercapto group, hydroxy group, halogen atom, cyano group, sulfonylgroup, an optionally substituted alkylthio group, an optionallysubstituted arylthio group, an optionally substituted alkylsulfinylgroup, an optionally substituted arylsulfinyl group, an optionallysubstituted sulfamoyl group, an optionally substituted alkoxyl group, anoptionally substituted aryloxy group, an optionally substituted acyloxygroup, an optionally substituted alkoxycarbonyloxy group, an optionallysubstituted carbamoyloxy group, an optionally substituted amino group,an optionally substituted acylamino group, an optionally substitutedalkoxycarbonylamino group, an optionally substituted ureido group, anoptionally substituted sulfonylamino group, an optionally substitutedsulfamoylamino group, a formyl group, an optionally substituted acylgroup or an optionally substituted silyl group.

An optionally substituted alkylsulfonyl group represented by Y includes,for example, a 3-dimethylamino-propane-1-sulfonyl group,3-piperidin-1-yl-propane-1-sulfonyl group, pyridin-3-yl-methanesulfonylgroup, dimethylcarbamoylmethyl group,tetrahydro-pyran-2-yl-methanesulfonyl group, and2-(2-methoxy-ethoxy)-ethanesulfonyl group, and among them, the3-dimethylamino-propane-1-sulfonyl group and3-piperidin-1-yl-propane-1-sulfonyl group are preferable.

A sulfonylamino group represented by Y includes, for example, amethanesulfonylamino group, ethanesulfonylamino group, andbenzenesulfonylamino group, and among them, the methanesulfonylaminogroup is preferable.

In the present invention, as a substituent represented by Y, a hydroxygroup, mercapto group, an optionally substituted alkylsulfonyl group orsulfonylamino group are preferable, and among them, the hydroxy group isespecially preferable.

Among the compounds represented by the general formula (1), the compoundrepresented by the general formula (1-Y′H) where the substituentrepresented by Y is Y′—H (Y′ is S: sulfur atom, O: Oxygen atom, or N:nitrogen atom, H represents hydrogen atom) is often described by thegeneral formula (1′-Y′H). (1-Y′H) and (1′-Y′H) are tautomers and thesame compounds.

In the aforementioned general formula (1), X can be: a mercapto group;hydroxy group; halogen atom; nitro group; cyano group; an optionallysubstituted alkyl group, an optionally substituted alkenyl group or anoptionally substituted alkynyl group; an optionally substitutedcarbocyclic or heterocyclic aryl group; an optionally substitutedalkylthio group; an optionally substituted arylthio group, an optionallysubstituted alkylsulfinyl group, an optionally substituted arylsulfinylgroup, an optionally substituted alkylsulfonyl group, an optionallysubstituted arylsulfonyl group, an optionally substituted sulfamoylgroup, an optionally substituted alkoxyl group, an optionallysubstituted aryloxy group, an optionally substituted acyloxy group, anoptionally substituted alkoxycarbonyloxy group, an optionallysubstituted carbamoyloxy group, an optionally substituted ornon-substituted amino group, an optionally substituted acylamino group,an optionally substituted alkoxycarbonylamino group, an optionallysubstituted ureido group, an optionally substituted sulfonylamino group,an optionally substituted sulfamoylamino group, a formyl group, anoptionally substituted acyl group, an optionally substituted carboxylgroup, an optionally substituted alkoxycarbonyl group, an optionallysubstituted carbamoyl group or an optionally substituted silyl group.

An optionally substituted alkyl group represented by X includes, forexample, a methyl group, ethyl group, isopropyl group, tert-butyl group,2,2-dimethylpropyl group, cyclopropyl group, N,N-dimethylaminomethylgroup, N,N-dimethylaminoethyl group, morpholinylmethyl group,piperidinylmethyl group, hydroxymethyl group, 1-hydroxyethyl group,2-hydroxyethyl group, 1-hydroxy-1-methyl-ethyl group, methoxyethylgroup, methoxymethyl group, benzyl group, 2-phenylethyl group, andpyridylmethyl group, and the ethyl group, isopropyl group, tert-butylgroup and 2,2-dimethylpropyl group are preferable.

An acyl group represented by X includes an acetyl group, propionylgroup, pivaloyl group, and benzoyl group, and among them, the acetylgroup is preferable.

A carbamoyl group represented by X includes a dimethylcarbamoyl group,1-piperidinecarbonyl group, and 4-morpholinecarbonyl group, and amongthem, the dimethylcarbamoyl group is preferable.

An alkenyl group represented by X that may have a substituent includes a1-alkenyl group, and 2-alkenyl group, and the 1-alkenyl group includes,for example, an ethenyl group, isopropenyl group, 3-hydroxy-1-propenylgroup, 2-acetyl-ethenyl group, and 2-phenyl-ethenyl group, and the2-alkenyl group includes, for example, an allyl group, and 2-butenylgroup.

An optionally substituted alkynyl group represented by X includes a1-alkynyl group and 2-alkynyl group, and the 1-alkynyl group includes,for example, an ethynyl group, 3,3-dimethyl-1-butynyl group,2-phenyl-ethynyl group, and 2-trimethylsilyl-1-ethynyl group, and the2-alkynyl group includes, for example, a 2-propynyl group, 2-butynylgroup, 3-phenyl-2-propynyl group, 4,4-dimethyl-2-pentynyl group, and3-trimethylsilyl-2-propynyl group, and the 2-propynyl group and2-butynyl group are preferable.

An optionally substituted carbocyclic aryl group represented by Xincludes, for example, a phenyl group, naphthyl group, chlorophenylgroup, and methoxyphenyl group.

An optionally substituted heterocyclic aromatic substituent representedby X includes, for example, a pyridyl group, quinolyl group, pyrimidinylgroup, and furyl group.

A 2,4-dihydroxyphenyl group may be substituted by one to three Xs at anyof the 3, 5, or 6 position, and may be monosubstitution, disubstitutionor trisubstitution. For example, a compound in which the 5 position ofthe 2,4-dihydroxyphenyl group is substituted by X means a5-monosubstituent represented by the following general formula (3).

wherein X, R and Y are the same as X, R and Y as the aforementionedgeneral formula (1).

A compound represented by the general formula (1-1) includes, forexample, a compound represented by the following general formula (3-1),

wherein R and Y are the same as R and Y as the general formula (1)described above. X¹, X² and X³ are each independently the same meaningas X of the general formula (1) described above, orthe following general formula (3-2),

wherein R and Y are the same as R and Y as the general formula (1)described above. X⁴ is the same meaning as X of the general formula (1)described above.

In the general formula (3-1) and (3-2) described above, X¹ and X² may bethe same or different and are preferably a hydrogen atom, methyl groupor 1-propynyl group. X³ and X⁴ are preferably a hydrogen atom or methylgroup.

As a substituent represented by X of the general formula (1) describedabove, a chlorine atom, ethyl group, isopropyl group, tert-butyl group,2,2-dimethylpropyl group, 2-propynyl group or 2-butynyl group isespecially preferable.

As a compound represented by the general formula (1), among them, acompound represented by the following general formula (4),

wherein X represents a chlorine atom, ethyl group, isopropyl group,tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group or2-butynyl group; Y represents a mercapto group, an optionallysubstituted alkylsulfonyl group or hydroxy group; m is 0 or 1; and Arepresents a cyclic amino group, a compound represented by the followinggeneral formula (1-2),

wherein X represents a chlorine atom, ethyl group, isopropyl group,tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group or2-butynyl group; and Ar^(a) represents a 4-methoxyphenyl group,3-methoxyphenyl group, 3,4-dimethoxyphenyl group, 3,4,5-trimethoxyphenylgroup or 3,4-methylenedioxyphenyl group, and a compound represented bythe following general formula (1-3),

wherein X represents a chlorine atom, ethyl group, isopropyl group,tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group or2-butynyl group; and Alk represents an optionally substituted alkylgroup, are preferable.

Particular examples of the compound represented by the general formula(1) are shown in Table 1 to Table 3-2.

The prodrug of the present invention includes, for example, a compoundrepresented by the following general formula (5),

wherein OR¹ and OR² represent a substituent which is converted to ahydroxy group easily due to dissociation of O—R¹ bond and O—R² bond invivo; and any one of R¹ and R² may be a hydrogen atom. R¹ and R²include, for example: an acyl group such as an acetyl group, andtrifluoroacetyl group; a carbamoyl group such as a dimethylcarbamoylgroup; an alkoxycarbonyl group such as a methoxycarbonyl group; aphosphoryl group such as a (MeO)₂P(═O)—; and an alkoxymethyl group suchas a methoxymethyl group.

The triazole derivative of the present invention may form a salt with anacid or a base, and the present invention includes a HSP90 inhibitor anda cancer therapeutic agent containing a salt of a compound representedby the general formula (1) as an active ingredient. A salt with an acidincludes, for example, an inorganic acid salt such as a hydrochloride,hydrobromide, and sulfate and a salt with an organic acid such as atrifluoroacetic acid, methanesulfonic acid, and p-toluenesulfonic acid.A salt with a base includes, for example, a sodium salt. These salts canbe produced by a standard method, and particular examples include forexample, the following compounds.

-   4-{5-Hydroxy-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol    (OH-a01) hydrochloride,-   4-{5-hydroxy-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol    (OH-a02) hydrochloride,-   5-[5-(but-2-ynyl)-2,4-dihydroxy-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-c02) hydrochloride,-   5-[2,4-dihydroxy-5-(prop-2-ynyl)-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-e02) hydrochloride,-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a13) dihydrochloride,-   5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[2-(morpholin-4-yl)-pyrimidin-5-yl]-2,4-dihydro-[1,2,4]triazol-3-one    (OH-a17) hydrochloride,-   4-isopropyl-6-{5-methanesulfonyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    (SFN-a02) hydrochloride,-   4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol    (SFN2-a08) hydrochloride,-   4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(3-piperidin-1-yl-propane-1-sulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol    (SFN3-a08) hydrochloride,-   4-(but-2-ynyl)-6-{5-mercapto-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    trifluoroacetate (SH-c02-TF),-   4-isopropyl-6-{5-methylsulfanyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    trifluoroacetate (SMe-a02-TF),-   4-isopropyl-6-{5-methylsulfinyl-4-[4-(morpholin-4-oxide-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    trifluoroacetate (SFX-a07-TF), and-   4-bromo-6-{5-methylsulfanyl-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol    trifluoroacetate (SMe-d01-TF).

The compound of the present invention can be produced, for example, inthe following way.

In the scheme (1), X and R have the same meaning as X and R in thegeneral formula (1). Hal represents a halogen atom. Pro represents aprotective group of a hydroxy group. Z represents an oxygen atom orsulfur atom. R³ represents an alkyl group. R⁴ represents an alkyl groupor aryl group. Each step will be described below.

Step A: a step for halogenating a resorcinol derivative represented bythe general formula (IMO). The halogen atom includes a chlorine atom,bromine atom or iodine atom, and among them, the bromine atom ispreferable. In the case where the halogen atom is the bromine atom, abrominating agent includes N-bromosuccinimide, benzyltrimethylammoniumtribromide, and bromine, and among them benzyltrimethylammoniumtribromide is preferable. In this step, it is preferable to usebenzyltrimethylammonium tribromide as a halogenating agent, and thereaction is carried out in a halogenated solvent at a temperature from0° C. to 50° C.

Step B: a step for protecting the hydroxy group of the resorcinolderivative represented by the general formula (IM1). A protective group,Pro, that may be used in this step includes, for example, analkoxymethyl group, substituted or non-substituted benzyl group, andsilyl group. Among them, the alkoxymethyl group such as a methoxymethylgroup, and benzyloxymethyl group are preferable, and the methoxymethylgroup is especially preferable. In the case where Pro is themethoxymethyl group, for example, methoxymethylchloride can be used as amethoxymethylating agent, and the reaction can be carried out in asolvent such as a polar non-proton solvent such as dimethylformamide, ahalogenated solvent, a nitrile solvent such as acetonitrile or an ethersolvent, in the presence of a base such as triethylamine, pyridine,diisopropylethylamine or potassium carbonate, at a temperature from −20°C. to 60° C.

Step C: a step for exchanging the halogen atom in the halogensubstituted resorcinol derivative represented by the general formula(IM2) with a metal atom such as lithium, and then converting to acarboxy group. The halogen-lithium exchange reaction is carried out, forexample, in an ether solvent such as tetrahydrofuran, and diethylether,using n-butyllithium at a temperature from −100° C. to 0° C., preferablyfrom −60° C. to −30° C. Then dry ice was added to the mixture, and thereaction was performed at a temperature from −80° C. to 50° C. to afforda carboxylic acid (IM3).

Step D: a step for synthesizing an acylhydrazide derivative representedby the general formula (IM4) by reacting a carboxylic acid derivativerepresented by the general formula (IM3) with hydrazine. The presentstep can be carried out, for example, using carbonyldiimidazole or thelike in tetrahydrofuran solution by adding benzylbromide or the like asnecessary to produce 1-acyl-3-benzylimidazolium salt, and then reactingwith hydrazine. Alternatively, the reaction with hydrazine can becarried out, using a carbodiimide such as dicyclohexylcarbodiimide or1-(3-dimethylaminopropyl)-3-ethylcarbodiimide as a condensation agent,and under coexistence of an activating agent such as1-hydroxybenzotriazole as necessary, in a solvent such asdimethylformamide.

A compound represented by the general formula (IM5-S) and generalformula (IM5-O) can be synthesized from an acylhydrazide derivativerepresented by the general formula (IM4) by Step E or Step G, or from acarboxylic acid derivative represented by the general formula (IM3) byStep F.

Step E: a step for synthesizing a derivative represented by the generalformula (IM5-S) or general formula (IM5-O) by reacting an acylhydrazidederivative represented by the general formula (IM4) with anisothiocyanate derivative represented by the general formula (IM8,Z=sulfur atom) or an isocyanate derivative represented by the generalformula (IM8, Z=oxygen atom). The present step can be carried out, forexample, by reacting an acylhydrazide derivative and isothiocyanate orisocyanate at a temperature from 0° C. to 150° C., preferably from 50°C. to 100° C. in a solvent such as ethanol, tert-butanol, anddimethylformamide.

Step F: a step for synthesizing a derivative represented by the generalformula (IM5) by reacting a carboxylic acid derivative represented bythe general formula (IM3) and a compound represented by the generalformula (IM9). The present step can be carried out, for example, byreacting in a solvent such as dimethylformamide, tetrahydrofuran, andN-methylpyrrolidone, using a condensation agent, for example,dicyclohexylcarbodiimide or1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, under the coexistence ofan activating agent such as 1-hydroxybenzotriazole as necessary, at atemperature from −20° C. to 50° C., preferably from 0° C. to 30° C.

Step G: a step for synthesizing a derivative represented by the generalformula (IM5-S) or general formula (IM5-O) by reacting an acylhydrazidederivative represented by the general formula (IM4) with a thiocarbamatederivative or carbamate derivative represented by the general formula(IM10). The present step can be carried out, for example, by reacting anacylhydrazide derivative with a thiocarbamate derivative or a carbamatederivative represented by the general formula (IM10) in a solvent suchas ethanol, tert-butanol, and dimethylformamide at a temperature from 0°C. to 150° C., preferably 50° C. to 100° C.

A compound represented by the general formula (IM6-S) or general formula(IM6-O) can be synthesized from a compound represented by the generalformula (IM5-S) or general formula (IM5-O) by Step H.

Step H: a step for synthesizing a triazole derivative represented by thegeneral formula (IM6-S) or (IM6-O) by a ring closure reaction of acompound represented by the general formula (IM5-S) or general formula(IM5-O). The present step can be carried out, for example, by reactingin the presence of a base such as sodium hydroxide, and potassiumhydroxide in a solvent such as water, and ethanol at a temperature from20° C. to 150° C., preferably from 70° C. to 120° C.

Step I: a step for synthesizing an alkylsulfanyltriazole derivativerepresented by the general formula (IM7-SR3) by alkylating atriazolethione derivative represented by the general formula (IM6-S)using an alkylating agent. The alkylating agent includes an alkylhalidesuch as methyl iodide, and a sulfonic acid alkyl ester or the like. Thepresent step can be carried out, for example, by reacting atriazolethione derivative represented by the general formula (IM6-S) andan alkyl halide such as methyl iodide, in a solvent such as an ethersolvent such as tetrahydrofuran, and diethyl ether, a polar solvent suchas dimethylformamide, a halogen solvent such as dichloromethane, or ahydrocarbon solvent such as toluene.

Step J and Step K: steps for deprotecting the protective group of thehydroxy group of a compound in which the hydroxy group is protected,represented by the general formulas (IM6-S), (IM6-O) or (IM7-SR3), andfor producing a benzene 1,3-diol derivative represented by the generalformulas (1) —SH, (1) —OH or (1) —SR3. In the case where the protectivegroup of the hydroxy group is a methoxymethyl group, the steps can becarried out under an acidic condition. Any of catalysts, which are knownto deprotect a methoxymethyl group and have no effect other than theprotective group, may be used as an acid catalyst which includes: aninorganic acid such as hydrochloric acid, and sulfuric acid; a sulfonicacid such as toluenesulfonic acid, methanesulfonic acid, andtrifluorosulfonic acid; a carboxylic acid such as acetic acid, andtrifluoroacetic acid; and a strong acid-weak base salt such aspyridiniumparatoluenesulfonate. In the case where the protective groupis a methoxymethyl group, it is preferable to carry out the reaction,for example, using 0.5-5.0 N hydrochloric acid as an acid catalyst, in amixed solvent of water and ethanol, methanol, tetrahydrofuran or thelike, at a temperature from 10° C. to 40° C., with a reaction time for 3hours to 3 days.

The isocyanate derivative (IM8:Z=oxygen atom) used in Step E can besynthesized, for example, according to the method described in Angew.Chem. Int. Ed. Engl. 26, 894 (1987).

The isothiocyanate derivative (IM8:Z=sulfur atom) used in Step E can besynthesized, for example, according to the method described inWO9921845.

The derivative (IM9) used in Step F and the carbamate derivative (IM10)used in Step G can be synthesized, for example, according to the methoddescribed in Chem. Pharm. Bull. 48 (12) 1935-1946 (2000).

When the triazole derivative of the present invention, a prodrug thereofor a pharmaceutically acceptable salt thereof is used as an anticanceragent, it can be administered safely, orally or parenterally (systemicadministration, local administration or the like), in a formulation suchas powder, granules, tablets, caplets, capsules, injectables,suppositories, and ointments, singly or mixed with pharmaceuticallyacceptable additives such as carriers, excipients, disintegrators,binders, lubricants, fluidizers, coating agents, suspending agents,emulsifiers, stabilizers, preservatives, flavoring agents, aroma agents,diluents, and solubilizers. The content of the triazole derivative ofthe present invention, a prodrug thereof or a pharmaceuticallyacceptable salt thereof in a formulation may be different depending onthe formulation but normally, it is preferable to be 0.1-100 weight %.The dosage may be different depending on the administration route, theage of a patient, the actual symptom to be prevented or treated and thelike, but, for example, in the case where the drug is orallyadministered to an adult, the dosage can be 0.01 mg to 2000 mg as anactive ingredient per day, preferably 0.1 mg to 1000 mg, which can beadministered once a day or several times a day.

The triazole derivative of the present invention, a prodrug thereof or apharmaceutically acceptable salt thereof possess a HSP90 inhibitoryactivity and are useful as the cancer therapeutic agent.

EXAMPLES

Next, the present invention will be described more particularly by theExamples but the present invention is not limited at all by theseexamples. In addition, the pharmacological test results of therepresentative compounds of the present invention demonstrating theefficacy of the compound of the present invention are shown in Table 4-1to 6-4.

The analytical method of LC/MS for the compounds of Examples are asfollows:

1)Instrument: Shimadzu LCMS-QP8000 alpha

Column: Inertsil ODS-III, 2.1 mm i.d.×100 mm,

Mobile phase A: acetonitrile/formic acid (99.9/0.1)Mobile phase B: water/formic acid (99.9/0.1)

Gradient:

Time (minute) 0.0 5.5 5.51 10.0 A Concentration a 100 a aFlow rate: 0.3 mL/minuteMethod 1) a=20; Method 2) a=52)

Instrument: Shimadzu LCMS-2010A Column: Inertsil ODS-III, 2.1 mmi.d.×100 mm,

Mobile phase A: acetonitrile/formic acid (99.9/0.1)Mobile phase B: water/formic acid (99.9/0.1)

Gradient:

Time (minute) 0.0 5.5 6.5 6.51 10.0 A concentration a 90 90 a aFlow rate: 0.3 mL/minuteMethod 3) a=20; Method 4) a=5; Method 5) a=40; Method 6) a=0; Method 7)a=60.

Example 1-1 Preparation of4-isopropyl-6-{5-mercapto-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SH-a01) and trifluoroacetate thereof (SH-a01-TF)

The First Step: Preparation of 4-bromo-6-isopropyl-benzene-1,3-diol(IM1-a)

4-isopropyl-benzene-1,3-diol (IMO-a: 9.13 g, 60 mmol) andbenzyltrimethyl ammoniumtribromide (24.6 g, 63 mmol) and methylenechloride 250 mL were placed in a 500 mL three-neck flask and werestirred at room temperature for 4 hours. After the reaction wasfinished, the reaction mixture was washed twice with saturated ammoniumchloride and once with saturated sodium chloride, and then dried overanhydrous sodium sulfate. After filtration and evaporation, the residuewas purified by silica gel column chromatography to obtain the titlecompound (IM1-a: 10.5 g, 76%).

LC/MS (Method 3): m/z (ESI, POS): 229, 231 [M+H]⁺; retention time: 5.68minutes.

¹H-NMR (200 MHz, CDCl₃, TMS) ppm: 7.21 (1H, s), 6.48 (1H, s), 5.33 (1H,s), 4.87 (1H, s), 3.10 (1H, sept, J=6.9 Hz), 1.22 (6H, d, J=6.9 Hz).

The material compound, 4-isopropyl-benzene-1,3-diol (IMO-a) wassynthesized by the method according to WO 04/72051 (Patent Document 6).

The Second Step: Preparation of1-bromo-5-isopropyl-2,4-bis-methoxymethoxy-benzene (IM2-a)

4-bromo-6-isopropyl-benzene-1,3-diol (IM1-a: 10.5 g, 45.5 mmol),dimethyl formamide (50 mL) and ethyldiisopropylamine (39.6 mL, 227 mmol)were placed in a 100 mL eggplant shaped flask, and the solution wascooled to 0° C., mixed with methoxymethylchloride (17.3 mL, 227 mmol)and the mixture was allowed to come to room temperature. After stirringfor 12 hours the solution was heated to 50° C. and stirring wascontinued further for 6.5 hours. After completing the reaction, ethylacetate was added, and after washing the organic layer 4 times withsaturated sodium chloride, the organic layer was dried anhydrous sodiumsulfate. After filtration and evaporation, the residue was purified bysilica gel column chromatography to obtain the title compound (IM2-a:12.1 g, 83.1%).

LC/MS (Method 3): m/z (ESI, POS): 318, 320 [M+H]⁺; retention time: 7.72minutes.

¹H-NMR (200 MHz, CDCl₃, TMS) ppm: 7.32 (1H, s), 6.93 (1H, s), 5.21 (2H,s), 5.18 (2H, s), 3.53 (3H, s), 3.49 (3H, s), 3.24 (1H, sept, J=6.8 Hz),1.19 (6H, d, J=6.8 Hz).

The Third Step: Preparation of5-isopropyl-2,4-bis-methoxymethoxy-benzoic acid (IM3-a)

1-bromo-5-isopropyl-2,4-bis-methoxymethoxy-benzene (IM2-a: 12.0 g, 37.8mmol) and tetrahydrofuran (150 mL) were placed in a 300 mL three-neckflask and cooled to −60° C. To this mixture a hexane solution ofN-butyllithium (24 mL, 1.59 M) was added slowly, and after adjusting thetemperature of the solution to −40° C. the mixture was stirred for 1hour. Dry ice powder was added to the reaction mixture and the reactionmixture was allowed to come to room temperature. After completing thereaction, the reaction mixture was mixed with distilled water andextracted with ethyl acetate. The extracted organic layer was dried overanhydrous sodium sulfate. After filtration and evaporation, the solidmaterial obtained was purified by suspending in hexane to obtain thetitle compound (IM3-a: 6.6 g, 62%).

LC/MS (Method 3): m/z (ESI, POS): 285 [M+H]⁺; retention time: 5.83minutes.

¹H-NMR (200 MHz, CDCl₃, TMS) ppm: 10.6 (1H, brs), 8.03 (1H, s), 7.27(1H, s), 5.40 (2H, s), 5.27 (2H, s), 3.57 (3H, s), 3.50 (3H, s), 3.26(1H, sept, J=6.9 Hz), 1.23 (6H, d, J=6.9 Hz).

The Fourth Step: Preparation of5-isopropyl-2,4-bis-methoxymethoxy-benzoic acid hydrazide (IM4-a)

5-isopropyl-2,4-bis-methoxymethoxy-benzoic acid (IM3-a: 2.84 g, 10mmol), N,N′-carbonyldiimidazole (1.62 g, 10 mmol) and tetrahydrofuran100 mL were placed in a 300 mL eggplant shaped flask and stirred at roomtemperature for 4 hours. The reaction mixture was mixed withbenzylbromide (1.19 mL, 10 mL), stirred at room temperature for further4 hours and then mixed with hydrazine mono hydrate (0.63 mL, 13 mmol)and stirred at room temperature overnight. After completing thereaction, most of the reaction mixture was removed under reducedpressure, and the residue was mixed with ethyl acetate. The organiclayer was washed with saturated sodium hydrogen carbonate and saturatedsodium chloride solutions, dried over anhydrous sodium sulfate. Afterfiltration and evaporation, the residue was purified by silica gelcolumn chromatography to obtain the title compound (IM4-a: 2.44 g, 82%).

LC/MS (Method 3): m/z (ESI, POS): 299 [M+H]⁺; retention time: 4.54minutes.

¹H-NMR (200 MHz, CDCl₃, TMS) ppm: 8.85 (1H, brs), 8.05 (1H, s), 6.93(1H, s), 5.30 (2H, s), 5.24 (2H, s), 3.52 (3H, s), 3.50 (3H, s), 3.26(1H, sept, J=6.9 Hz), 1.23 (6H, d, J=6.9 Hz).

The Fifth Step: Preparation of4-[4-(morpholin-4-yl)-phenyl]-1-(5-isopropyl-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide (IM5-S-a01)

5-isopropyl-2,4-bis-methoxymethoxy-benzoic acid hydrazide (IM4-a: 29.8mg, 0.1 mmol), 4-(4-isothiocyanate-phenyl)-morpholine (23.4 mg, 0.1mmol) and ethanol (1 mL) were placed in a test tube and heated underreflux for 2 hours. After completing the reaction the reaction mixturewas concentrated under reduced pressure to obtain crude crystals of4-(4-morpholin-4-yl-phenyl)-1-(5-isopropyl-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide(IM5-S-a01). These crude crystals were subjected to the next reactionwithout purification in particular.

LC/MS (Method 1): m/z (ESI, POS): 519 [M+H]⁺; retention time: 6.54minutes.

The Sixth Step: Preparation of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-yl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-a01)

Crude crystals of4-[4-(morpholin-4-yl)-phenyl]-1-(5-isopropyl-2,4-bis-methoxymethoxy-benzoyl)thiosemicarbazide(IM5-S-a01) and 5% aqueous sodium hydroxide (1 mL) were placed in a testtube and refluxed for 2 hours. After completing the reaction, thereaction mixture was extracted with methylene chloride, and the organiclayer was combined drying over anhydrous sodium sulfate. Filtration andevaporation gave crude crystals of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-morpholin-4-yl-phenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-a01). These crude crystals were subjected to the next reactionwithout purification in particular.

LC/MS (Method 1): m/z (ESI, NEG): 499 [M+H]⁻; retention time: 6.44minutes.

The Seventh Step: Preparation of4-isopropyl-6-{5-mercapto-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SH-a01) and trifluoroacetate salt thereof (SH-a01-TF)

Crude crystals of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-yl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-a01) and a mixed solvent of 5.0 N aqueous hydrochloric acid (1mL) and ethanol (1 mL) were placed in a test tube and stirred at roomtemperature for 24 hours. After completing the reaction, the reactionmixture was neutralized with 10 N aqueous sodium hydroxide solution andthen extracted with ethyl acetate and methylene chloride, and thecollected organic layer was dried over anhydrous sodium sulfate. Afterfiltration and evaporation, thus obtained residue was purified by HPLCfractionation to obtain the title compound (SH-a01: 7.6 mg, 17%: 3 stepsfrom IM4-a).

LC/MS (Method 1): m/z (ESI, POS): 413 [M+H]⁺; retention time: 5.51minutes.

Also, by using 0.1% trifluoroacetic acid-acetonitrile/water in thepurification step by HPLC fractionation, the trifluoroacetate salt wassimilarly obtained (SH-a01-TF).

Example 1-2 Preparation of4-isopropyl-6-{5-mercapto-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SH-a02)

The First Step: Preparation of4-[4-(morpholin-4-ylmethyl)-phenyl]-1-(5-isopropyl-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide(IM5-S-a02)

5-isopropyl-2,4-bis-methoxymethoxy-benzoic acid hydrazide (IM4-a: 1.99g, 6.67 mmol), 4-(4-isothiocyanate-benzyl)-morpholine (1.72 g, 7.34mmol) and ethanol (30 mL) were placed in a 100 mL eggplant shaped flaskand heated under reflux for 2 hours. After completing the reaction, thereaction mixture was concentrated under reduced pressure to obtain4-[4-(morpholin-4-ylmethyl)-phenyl]-1-(5-isopropyl-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide(IM5-S-a02). This crude product was subjected to the next reactionwithout purification in particular.

LC/MS (Method 3): m/z (ESI, POS): 533 [M+H]⁺; retention time: 3.72minutes.

The Second Step: Preparation of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-a02)

Crude crystals of4-[4-(morpholin-4-ylmethyl)-phenyl]-1-(5-isopropyl-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide(IM5-S-a02) and 5% aqueous sodium hydroxide (30 mL) were placed in a 100mL eggplant shaped flask and refluxed for 2 hours. After completing thereaction, the reaction mixture was extracted with methylene chloride,and the organic layer was combined, dried over anhydrous sodium sulfate.After filtration and evaporation, the residue thus obtained was purifiedby silica gel column chromatography to obtain the title compound(IM6-S-a02: 1.43 g, 41.6%).

LC/MS (Method 3): m/z (ESI, POS): 515 [M+H]⁺; retention time: 3.42minutes.

The Third Step: Preparation of4-isopropyl-6-{5-mercapto-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SH-a02)

Crystals of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-a02) (1.43 g, 2.77 mmol) and a mixed solvent of 5.0 N aqueoushydrochloric acid (15 mL) and ethanol (15 mL) were placed in a 100 mLeggplant shaped flask and stirred at room temperature for 3.5 hours.After completing the reaction, the reaction mixture was neutralized with5 N aqueous sodium hydroxide and then extracted with ethyl acetate.After drying over anhydrous sodium sulfate the collected organic layerwas concentrated under reduced pressure. The residue thus obtained waspurified by silica gel column chromatography to obtain the titlecompound (SH-a02: 647 mg, 54.7%).

LC/MS (Method 4): m/z (ESI, POS): 427 [M+H]⁺; retention time: 3.83minutes.

¹H-NMR (200 MHz, DMSO-d₆, TMS) ppm: 13.9 (1H, s), 9.62 (1H, s), 9.43(1H, s), 7.32 (2H, d, J=8.1 Hz), 7.18 (2H, d, J=8.1 Hz), 6.80 (1H, s),6.23 (1H, s), 3.62-3.52 (4H, m), 3.45 (2H, s), 2.94 (1H, sept, J=6.8Hz), 2.40-2.25 (4H, m), 0.94 (6H, d, J=6.8 Hz).

Example 1-3 Preparation of4-[4-(4-bromo-phenyl)-5-mercapto-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SH-a03)

The title compound (SH-a03) was synthesized in a similar manner asdescribed in Example 1-1.

LC/MS (Method 5): m/z (ESI, POS): 406, 408 [M+H]⁺; retention time: 4.38minutes.

¹H-NMR (200 MHz, CDCl₃, TMS) ppm: 9.25 (1H, s), 7.75 (2H, d, J=8.7 Hz),7.26 (2H, d, J=8.7 Hz), 6.43 (1H, s), 6.42 (1H, s), 2.91 (1H, sept,J=6.9 Hz), 0.81 (6H, d, J=6.9 Hz).

Example 1-4 Preparation of4-(but-2-ynyl)-6-{5-mercapto-4-[4-morpholin-4-ylmethyl]-phenyl}-4H-[1,2,4]trioazol-3-yl]-benzene-1,3-dioltrifluoroacetate (SH-c02-TF)

The First Step: Preparation of 2,4-bis-aryloxy-5-bromo-benzoic acidallyl ester (IMO2-c)

5-bromo-2,4-dihydroxy-benzoic acid (IM01-c: 6.99 g, 30 mmol), potassiumcarbonate (16.58 g, 120 mmol) and dimethylformamide (60 mL) were placedin a 500 mL flask under a atmosphere of nitrogen, and while stirring atroom temperature, allylbromide (8.6 mL, 100 mmol) was added dropwise.After stirring at room temperature for 3 hours, the reaction mixture wasmixed with water (600 mL) and extracted twice with ethyl acetate (600mL). After washing with saturated sodium chloride solution, the organiclayer was dried over anhydrous sodium sulfate. Filtration andevaporation gave the title compound (IMO2-c: 10.4 g, 98%).

LC/MS (Method 3): m/z (ESI, POS): 355 [M+2+H]⁺; retention time: 7.79minutes.

The Second Step: Preparation of(2,4-bis-allyloxy-5-bromo-phenyl)methanol (IM03-c)

IM02-c (1.41 g, 4 mmol) and dichloromethane (20 mL) were placed in a 300mL three-neck flask under a atmosphere of nitrogen, cooled to −78° C.,and a toluene solution of 1.01M diisobutylaluminum hydride (8.8 mL) wasslowly added dropwise so that the inside temperature did not rise to−70° C. or higher. After stirring for 1 hour, methanol (4 mL) andsaturated ammonium chloride solution (20 mL) were added, and the mixturewas allowed to come to room temperature. The mixture was extracted twicewith chloroform (50 mL) (at this time 2M hydrochloric acid was addeduntil the phases were separated). After washing with saturated sodiumchloride solution, the organic layer was dried over anhydrous sodiumsulfate. After filtration and evaporation, the residue was purified withsilica gel column chromatography (hexane-ethyl acetate=2:1) to obtainthe title compound (IM03-c: 1.07 g, 90%).

LC/MS (Method 3): m/z (ESI, POS): 299 [M+H]⁺; retention time: 6.3minutes.

¹H-NMR (400 MHz, CDCl₃, TMS) ppm: 7.42 (1H,$), 6.23 (1H, s), 5.97-6.10(2H, m), 5.30-5.51 (4H, m), 4.61 (2H, s), 4.60-4.55 (4H, m).

The Third Step: Preparation of 2,4-bis-allyloxy-5-bromo-benzaldehyde(IM04-c)

IM03-c (0.55 g, 1.83 mmol), manganese dioxide (5.2 g, 59.8 mmol) andchloroform (50 mL) were placed in a 300 mL flask and stirred at roomtemperature for 72 hours. After filtering the reaction mixture, thefiltrate and wash were collected and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(hexane-ethyl acetate=3:1-2:1) to obtain the title compound (IM04-c:0.42 g, 78%).

LC/MS (Method 3): m/z (ESI, POS): 299 [M+2+H]⁺; retention time: 7.2minutes.

¹H-NMR (400 MHz, CDCl₃, TMS) ppm: 10.29 (1H, s), 8.02 (1H, s), 6.43 (1H,s), 6.11-6.07 (2H, m), 5.54-5.34 (4H, m), 4.66 (4H, m).

The Fourth Step: Preparation of1-(2,4-bis-allyloxy-5-bromo-phenyl)-but-2-ynyl-1-ol (IM05-c)

IM04-c (486 mg, 1.6 mmol) and anhydrous tetrahydrofuran (5 mL) wereplaced in a 30 mL two-neck flask under a nitrogen atmosphere, and atetrahydrofuran solution of 0.5M 1-propynylmagnesium bromide (8.0 mL,4.0 mmol) was added dropwise at 0° C. After stirring for 1 hour, thereaction mixture was mixed with saturated ammonium chloride solution (4mL) and saturated sodium chloride solution (10 mL), and extracted twicewith ethyl ether (20 mL). After washing with saturated sodium chloridesolution and drying over anhydrous sodium sulfate, filtration andevaporation gave the crude product of the title compound (IM05-c: 0.55g).

LC/MS (Method 3): m/z (ESI, POS): 321 [M+2+H—H₂O]⁺; retention time: 6.79minutes.

The Fifth Step: Preparation of1,5-bis-allyloxy-2-bromo-4-(but-2-ynyl)-benzene (IM06-c)

Crude IM05-c (0.55 g) dissolved in anhydrous acetonitrile (4 mL) wasplaced in a 50 mL two-neck flask under a nitrogen atmosphere, mixed withtriethylsilane (0.28 mL, 1.76 mmol) and boron trifluoride diethyl ethercomplex (0.223 mL, 1.76 mmol) under ice cold condition and stirred for 1hour. Potassium carbonate (553 mg, 4 mmol) and water (70 mL) were addedto the reaction mixture, which was extracted twice with ethyl acetate(70 mL). The organic layer was washed with saturated sodium chloridesolution, dried over anhydrous sodium sulfate. Filtration andevaporation gave the crude title compound (IM06-c: 0.51 g).

LC/MS (Method 3): m/z (ESI, POS): 321 [M+H]⁺; retention time: 8.22minutes.

The Sixth Step: Preparation of 2,4-bis-allyloxy-5-(but-2-ynyl)-benzoicacid (IM07-c)

Crude IM06-c (0.51 g) dissolved in anhydrous tetrahydrofuran (6 mL) wasplaced in a 50 mL two-neck flask under a nitrogen atmosphere, and aftercooling to −78° C., a hexane solution of 1.59M n-butyllithium (1.1 mL,1.76 mmol) was added dropwise. The reaction mixture was stirred for 1hour, mixed with dry ice (7 g) and further stirred for 1 hour. Thereaction mixture was adjusted to pH 2.5 by mixing with 10% potassiumhydrogensulfate and extracted twice with ethyl acetate (40 mL). Theorganic layer was washed with saturated sodium chloride solution, driedover anhydrous sodium sulfate. After filtration and evaporation, theresidue was purified by silica gel column chromatography(hexane-ethylacetate=2:1) to obtain the title compound (IM07-c: 0.28 g,yield 62%, 3 steps).

LC/MS (Method 3): m/z (ESI, POS): 287 [M+H]⁺; retention time: 6.52minutes.

¹H-NMR (400 MHz, CDCl₃, TMS) ppm: 10.63 (1H, brs), 8.26 (1H, s), 6.48(1H, s), 6.14-5.98 (2H, m), 5.54-5.32 (4H, m), 4.76 (2H, m) 4.61 (2H,m), 3.46 (2H, m), 1.85 (3H, t, J=2.6 Hz).

The Seventh Step: Preparation of4-[4-(morpholin-4-ylmethyl)-phenyl]-1-[5-(but-2-ynyl)-2,4-bismethoxymethoxy-benzoyl]thiosemicarbazide(F461-IM01)

2,4-bis-allyloxy-5-(but-2-ynyl)-benzoic acid (IM07-c: 286 mg, 1 mmol),4-[4-(morpholin-4-ylmethyl)-phenyl]thiosemicarbazide (266 mg, 1 mmol),1-hydroxybenzo triazole monohydrate (135 mg, 1 mmol) anddimethylformamide (3 mL) were placed in a 100 mL eggplant shaped flask.The reaction mixture was cooled to 0° C., and a dimethylformamidesolution of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (230 mg, 1.2mmol) (2 mL) was added to the reaction mixture dropwise slowly. Aftercompleting the dropwise addition, the reaction mixture was stirred for 4hours and allowed to come to room temperature. After completing thereaction, the reaction mixture was extracted with ethyl acetate, and theorganic layer was washed with saturated sodium chloride solution andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography to obtain the title compound (F461-IM01: 307mg, 57.5%).

LC/MS (Method 3): m/z (ESI, POS): 535 [M+H]⁺; retention time: 4.06minutes.

The Eighth Step: Preparation of5-[5-(but-2-ynyl)-2,4-bis-methoxymethoxy-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(F461-IMO2)

4-[4-(morpholin-4-ylmethyl)-phenyl]-1-[5-(but-2-ynyl)-2,4-bismethoxymethoxy-benzoyl]thiosemicarbazide(F461-IM01: 71.3 mg, 0.133 mmol) and 5% aqueous sodium hydroxide (5 mL)were placed in a 30 mL eggplant shaped flask and heated under reflux for2.5 hours. After completing the reaction, the reaction mixture wasextracted with methylene chloride and concentrated under reducedpressure. The crude product (F461-IMO2) thus obtained was subjected tothe next reaction without purification in particular.

LC/MS (Method 3): m/z (ESI, POS): 517 [M+H]⁺; retention time: 3.85minutes.

The Ninth Step: Preparation of4-but-2-ynyl-6-{5-mercapto-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl]-benzene-1,3-dioltrifluoroacetate (SH-c02-TF)

Crude5-[5-(but-2-ynyl)-2,4-bis-methoxymethoxy-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(F461-IMO2) obtained in the eighth step, potassium carbonate (110 mg,0.8 mmol) and triphenylphosphinepalladium (7.7 mg, 0.0067 mmol) wereplaced in a 30 mL eggplant shaped flask and heated under reflux underargon atmosphere for 3.5 hours. After completing the reaction, thereaction mixture was neutralized with 1 N hydrochloric acid andextracted with ethyl acetate. The residue thus obtained was purified byHPLC fractionation to obtain the title compound (SH-c02-TF: 10.9 mg,18.7%: 2 steps).

LC/MS (Method 4): m/z (ESI, POS): 437 [M+H]⁺; retention time: 3.80minutes.

¹H-NMR (400 MHz, CD₃OD, TMS) ppm: 7.57 (2H, d, J=8.4 Hz), 7.44 (2H, d,J=8.4 Hz), 7.21 (1H, S), 6.17 (1H, S), 4.38 (2H,$), 4.12-3.96 (2H, br),3.80-3.60 (2H, br), 3.55-3.35 (2H, br), 3.28-3.20 (2H, m), 3.17-3.15(2H, br), 1.82 (3H, t, J=2.4 Hz).

Example 1-5 Preparation of4-bromo-6-{5-mercapto-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SH-d01)

The First Step: Preparation of 5-bromo-2,4-bis-methoxymethoxybenzoicacid methoxymethyl ester (IM3-d)

Sodium hydride (6 g, 150 mmol) and tetrahydrofuran (100 mL) were placedin a 500 mL four-neck flask and cooled to 0° C. A tetrahydrofuransolution (50 mL) of 5-bromo-2,4-dihydroxybenzoic acid monohydrate (12.6g, 50 mmol) was added to it slowly dropwise, and stirring was continuedfor 30 minutes while keeping the temperature at 0° C. After thatmethoxymethylchloride (12.4 mL, 165 mmol) diluted in tetrahydrofuran (30mL) was added slowly dropwise. After completing the instillation, thereaction mixture was allowed to come to room temperature and stirred for24 hours. After completing the reaction, saturated aqueous ammoniumchloride was added, and the reaction mixture was extracted with ethylacetate, dried over anhydrous sodium sulfate. After filtration andevaporation, the residue thus obtained was purified by silica gel columnchromatography to obtain the title compound (IM3-d: 6.95 g, 38%).

LC/MS (Method 3) retention time: 6.29 minutes.

¹H-NMR (400 MHz, CDCl₃, TMS) ppm: 8.11 (1H, s), 7.03 (1H, s), 5.44 (2H,s), 5.30 (2H, s), 5.26 (2H, s), 3.55 (3H, s), 3.53 (3H, s), 3.52 (3H,s).

The Second Step: Preparation of 5-bromo-2,4-bis-methoxymethoxybenzoicacid hydrazide (IM4-d)

5-bromo-2,4-bis-methoxymethoxybenzoic acid methoxymethylester (IM3-d:4.36 g, 11.9 mmol), ethanol (15 mL) and hydrazine monohydrate (0.96 mL,29.8 mmol) were placed in a 100 mL eggplant shaped flask and stirred at70° C. for 27 hours. After completing the reaction, the reaction mixturewas extracted with methylene chloride, and the organic layer was washedwith saturated sodium chloride solution and then concentrated underreduced pressure. The solid material thus obtained was suspended inhexane for purification to obtain the title compound (IM4-d: 1.8 g,46%).

LC/MS (Method 3): m/z (ESI, POS): 336 [M+H]⁺; retention time: 4.10minutes.

The Third Step: Preparation of4-[4-(morpholin-4-yl)-phenyl]-1-(5-bromo-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide (IM5-S-d01)

5-bromo-2,4-bis-methoxymethoxy-benzoic acid hydrazide (IM4-d: 670 mg, 2mmol), 4-(4-isothiocyanate-phenyl)-morpholine (441 mg, 2 mmol) andethanol (10 mL) were placed in a 50 mL eggplant shaped flask and heatedunder reflux for 2 hours. After completing the reaction, the reactionmixture was concentrated under reduced pressure to obtain crude crystalsof 4-[4-(morpholin-4-yl)-phenyl]-1-(5-bromo-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide (IM5-S-d01). These crudecrystals were subjected to the next reaction without purification inparticular.

LC/MS (Method 3): m/z (ESI, POS): 555 [M+H]⁺; retention time: 5.80minutes.

The Fourth Step: Preparation of5-(5-bromo-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-yl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-d01)

The crude crystals of 4-[4-(morpholin-4-yl)-phenyl]-1-(5-bromo-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide (IM5-S-d01) and 5% aqueoussodium hydroxide (1 mL) were added in a test tube and refluxed for 2hours. After completing the reaction, the reaction mixture was extractedwith methylene chloride, and the combined organic layer was dried overanhydrous sodium sulfate. Filtration and evaporation gave crude crystalsof5-(5-bromo-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-yl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-d01) (2 steps, 35.6%). These crude crystals were subjected to thenext reaction without purification in particular.

LC/MS (Method 3): m/z (ESI, POS): 537 [M+H]⁺; retention time: 4.3minutes.

The Fifth Step: Preparation of4-bromo-6-{5-mercapto-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SH-d01)

The crude crystals of5-(5-bromo-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-yl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-d01) and a mixed solvent of 5.0 N aqueous hydrochloric acid (1mL) and ethanol (1 mL) were placed in a test tube and stirred at roomtemperature for 24 hours. After completing the reaction, the reactionmixture was neutralized with 10 N aqueous sodium hydroxide, and thenextracted with ethyl acetate and methylene chloride. The collectedorganic layer was dried over anhydrous sodium sulfate. After filtrationand evaporation, the residue thus obtained was purified by

HPLC fractionation to obtain the title compound (SH-d01: 8.6 mg, 34%).

LC/MS (Method 2): m/z (ESI, NEG): 447 [M+H]; retention time: 4.51minutes.

Example 1-6 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-2,4-dihydro-[1,2,4]-triazol-3-thione(SH-a08)

The First Step: Preparation of 4-methoxyphenylthiosemicarbazide (F53-02)

4-methoxyphenylthioisocyanate (10 g, 60.5 mmol) and ethanol (18 mL) wereplaced in a 100 mL eggplant shaped flask, and then the reaction mixturewas cooled to 0° C., to which an ethanol solution (18 mL) of hydrazinemonohydrate (2.9 mL, 90.8 mmol) was added dropwise slowly. The reactionmixture was stirred for 4 hours while the temperature was raised to roomtemperature. After completing the reaction, the deposited solids werefiltered under reduced pressure, washed with hexane to obtain the titlecompound (F53-02: 11.3 g, 94.6%).

LC/MS (Method 1): m/z (ESI, POS): 198 [M+H]⁺; retention time: 3.59minutes.

The Second Step: Preparation of4-methoxyphenyl-1-[5-isopropyl-2,4-bis(methoxymethoxy)-benzoyl]thiosemicarbazide(F53-03)

5-isopropyl-2,4-bis(methoxymethoxy)-benzoic acid (IM3-a: 16.4 g, 54.5mmol), 4-methoxyphenylthiosemicarbazide (F53-02: 11.3 g, 57.3 mmol),dimethylformamide (150 mL) and 1-hydroxybenzotriazole monohydrate (8.11g, 60.0 mmol) were placed in a 500 mL eggplant shaped flask. Thereaction mixture was cooled to 0° C. and mixed with a suspension of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (11.5 g, 60 mmol) indimethylformamide (100 mL). The mixture was stirred for 4 hours andallowed to come to room temperature. After completing the reaction,saturated sodium chloride solution (500 mL) was added to the reactionmixture, and the mixture was extracted twice with ethyl acetate (500mL). The organic layer thus obtained was washed 4 times with saturatedsodium chloride solution (500 mL) and dried over anhydrous sodiumsulfate. After removing sodium sulfate by filtration, the solvent wasdistilled under reduced pressure, and the solids obtained were suspendedin hexane (1000 mL) for purification and collected by filtration. Thesolids thus obtained were dried under reduced pressure to obtain thetitle compound (F53-03: 21.9 g, 83.8%).

LC/MS (Method 3): m/z (ESI, POS): 464 [M+H]⁺; retention time: 6.46minutes.

The Third Step: Preparation of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-methoxy-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(F53-04)

4-methoxyphenyl-1-[5-isopropyl-2,4-bis(methoxymethoxy)-benzoyl]thiosemicarbazide(F53-03: 24.9 g, 53.7 mmol) and 5% aqueous sodium hydroxide (500 mL)were placed in a 1 L eggplant shaped flask and heated under reflux for 1hour. After completing the reaction, the reaction mixture wasneutralized with saturated aqueous ammonium chloride, and the solidsdeposited were collected by filtration, washed with distilled water andthen dried under reduced pressure. The crude product thus obtained waspurified by suspending in ethyl acetate/hexane and concentrating underreduced pressure to obtain the title compound (F53-04:20.5 g, 85.4%).

LC/MS (Method 3): m/z (ESI, POS): 446 [M+H]⁺; retention time: 6.33minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 7.19 (1H, s), 7.16 (2H, d, J=9.0Hz), 6.92 (2H, d, J=9.0 Hz), 6.75 (1H, s), 5.20 (2H, s), 4.94 (2H, s),3.73 (3H, s), 3.37 (3H, s), 3.21 (3H, s), 3.14 (1H, sept, J=6.8 Hz),1.07 (6H, d, J=6.8 Hz).

The Fourth Step: Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-2,4-dihydro-[1,2,4]-triazole-3-thione(SH-a08)

The title compound (SH-a08, 12 mg, 62.2%) was obtained by using5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-methoxy-phenyl]-2,4-dihydro[1,2,4]triazol-3-thione(F53-04, 24.1 mg, 0.054 mmol), in place of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-morpholin-4-ylmethyl)-phenyl]-2,4-dihydro[1,2,4]triazol-3-one,and treating in the similar manner as in the fourth step in the Example2-7.

LC/MS (Method 3): m/z (ESI, POS): 358 [M+H]⁺; retention time: 5.31minutes.

¹H-NMR (400 MHz, CD₃OD, TMS) ppm: 7.19 (2H, d, J=9.0 Hz), 6.99 (2H, d,J=9.0 Hz), 6.73 (1H, s), 6.26 (1H, s), 3.82 (3H, s), 3.04 (1H, sept,J=7.0 Hz), 0.94 (6H, d, J=7.0 Hz).

Example 1-7 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-phenyl-2,4-dihydro-[1,2,4]-triazol-3-one(SH-a15)

The title compound (SH-a15) was obtained by the similar process to thatof Example 1-6 in 4 steps using phenylisothiocyanate in place of F53-01of Example 1-6.

LC/MS (Method 3): m/z (ESI, POS): 328 [M+H]⁺; retention time: 5.27minutes.

Example 1-8 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-pyridin-3-yl-2,4-dihydro-[1,2,4]-triazol-3-one(SH-a16)

The title compound (SH-a16) was obtained by the similar process to thatof Example 1-6 in 4 steps using 3-isothiocyanate-pyridine in place ofF53-01 of Example 1-6.

LC/MS (Method 3): m/z (ESI, POS): 329 [M+H]⁺; retention time: 4.29minutes.

Example 1-9 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-isopropyl-2,4-dihydro-[1,2,4]-triazol-3-thione(SH-a21)

The title compound [SH-a21, 13.8 mg, 47% in 3 steps from IM4-a (thestarting material of Example 1-2)] was obtained by a similar process tothe fourth step of Example 2-7 using5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-isopropyl-2,4-dihydro-[1,2,4]triazol-3-thion(an intermediate of Example 2-13, F63-03) in place of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one.

LC/MS (Method 3): m/z (ESI, POS): 294 [M+H]⁺; retention time: 5.04minutes.

¹H-NMR (400 MHz, CD₃OD, TMS) ppm: 6.96 (1H, s), 6.41 (1H, s), 4.63 (1H,sept, J=7.0 Hz), 3.18 (1H, sept, J=6.8 Hz), 1.21 (6H, d, J=7.0 Hz) 1.18(6H, d, J=6.8 Hz)

Example 1-10 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-isobutyl-2,4-dihydro-[1,2,4]triazol-3-thione(SH-a22)

The bis(methoxymethyl) protected compound of the title compound (SH-a22)was obtained in 3 steps by a similar process to that of Example 2-13using isobutylisothiocyanete in place of isopropylisothiocyanate ofExample 2-13. This compound was deprotected by a similar operation tothat in the fourth step of Example 1-6 to obtain the title compound(SH-a22).

LC/MS (Method 3): m/z (ESI, POS): 308 [M+H]⁺; retention time: 5.39minutes.

Example 1-11 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-cyclohexyl-2,4-dihydro-[1,2,4]triazol-3-thione(SH-a23)

The bis(methoxymethyl) protected compound of the title compound (SH-a23)was obtained in 3 steps by a similar process to that of Example 2-13using cyclohexylisothiocyanate in place of isopropylisothiocyanate ofExample 2-13. This compound was deprotected by a similar operation tothat in the fourth step of Example 1-6 to obtain the title compound(SH-a23).

LC/MS (Method 3): m/z (ESI, POS): 334 [M+H]⁺; retention time: 5.71minutes.

¹H-NMR (400 MHz, CD₃OD, TMS) ppm: 6.95 (1H, s), 6.41 (1H, s), 4.30-4.20(1H, brs), 3.31 (1H, sept, J=7.0 Hz), 2.40-2.20 (2H, brs), 1.85-1.70(4H, brs), 1.66-1.55 (1H, brs), 1.35-1.20 (2H, m), 1.18 (6H, d, J=7.0Hz), 1.10-1.00 (1H, m).

Example 1-12 Preparation of4-{1-benzyl-piperidin-4-yl}-5-(2,4-dihydroxy-5-isopropyl-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(SH-a25) trifluoroacetate

The First Step: Preparation of4-(1-benzyl-piperidin-4-yl)-1-(5-isopropyl-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide(F66-02)

Tetrahydrofuran (5 mL), triethylamine (0.086 mL, 1.25 mmol),thiophosgene (0.042 mL, 0.55 mmol) and 4-amino-1-benzyl-piperidine(0.112 mL, 0.55 mmol) were placed in a 10 mL eggplant shaped flask andstirred at room temperature for 2 hours, and then5-isopropyl-2,4-bis-methoxymethoxy-benzoic acid hydrazide (IM4-a, 149mg, 0.5 mmol) was added. The reaction mixture was stirred at roomtemperature for 24 hours. After completing the reaction, the reactionmixture was concentrated under reduced pressure. The residue obtainedwas subjected to the next reaction without further purification.

LC/MS (Method 3): m/z (ESI, POS): 531 [M+H]⁺; retention time: 3.92minutes.

The Second Step: Preparation of4-(1-benzyl-piperidin-4-yl)-5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(F66-03)

4-(1-benzyl-piperidin-4-yl)-1-(5-isopropyl-2,4-bismethoxymethoxy-benzoyl)thiosemicarbazide(the crude product F66-02 of the previous step) and 5% aqueous sodiumhydroxide were placed in a 10 mL eggplant shaped flask and heated underreflux for 3 hours. After completing the reaction, the reaction mixturewas extracted with methylene chloride, and the extract was dried overanhydrows sodium sulfate. After filtration and evaporation, the residuethus obtained was subjected to the next reaction without furtherpurification.

LC/MS (Method 3): m/z (ESI, POS): 513 [M+H]⁺; retention time: 3.94minutes.

The Third Step: Preparation of4-(1-benzyl-piperidin-4-yl)-5-(2,4-dihydroxy-5-isopropyl-phenyl)-2,4-dihydro-1,2,4-triazol-3-one(SH-a25) trifluoroacetate

4-(1-benzyl-piperidin-4-yl)-5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(the crude product F66-03 of the previous step), ethanol (3 mL) and 5 Nhydrochloric acid (3 mL) were placed in a test tube and at roomtemperature for 24 hours. After completing the reaction, the reactionmixture was extracted with ethyl acetate, and the organic layer waswashed 4 times with saturated sodium chloride solution, dried overanhydrous sodium sulfate. After filtration and evaporation, the titlecompound (SH-a25 trifluoroacetate, 40 mg, 14.9%) was obtained bypurifying the residue by HPLC fractionation.

LC/MS (Method 3): m/z (ESI, POS): 425 [M+H]⁺; retention time: 3.35minutes.

Example 1-13 Preparation of5-(2,4-dihydroxy-5-isopropylphenyl)-4-(2-pyridin-3-ylethyl)-2,4-dihydro-[1,2,4]triazol-4-thione(SH-a28)

The bis(methoxymethyl) protected compound of the title compound (SH-a28)was obtained in 4 steps by a similar process to that of Example 2-2(B)using 3-(2-aminoethyl)pyridine in place of4-morpholin-4-ylmethyl-phenylamine (F45-000) of Example 2-2(B). Thiscompound was deprotected by a similar operation to that in the seventhstep of Example 2-13 to obtain the title compound (SH-a28).

LC/MS (Method 6): m/z (ESI, POS): 357 [M+H]⁺; retention time: 4.32minutes.

¹H-NMR [400 MHz, CDCl₃—CD₃OD (three drops)] δ 1.18 (d, J=7.0 Hz, 6H),3.10-3.23 (m, 3H), 4.25 (t, J=7.7 Hz, 2H), 6.39 (s, 1H), 6.95 (s, 1H),7.23 (dd, J=4.9, 7.9 Hz, 1H), 7.57 (d, J=7.9 Hz, 1H), 8.17 (s, 1H), 8.36(d, J=4.9 Hz, 1H).

Example 1-14 Preparation of5-(2,4-dihydroxy-5-isopropylphenyl)-4-(tetrahydrofuran-2-ylmethyl)-2,4-dihydro-[1,2,4]triazol-3-thione(SH-a31)

The bis(methoxymethyl) protected compound of the title compound (SH-a31)was obtained in 4 steps by a similar process to that of Example 2-2(B)using tetrahydrofurfurylamine in place of4-morpholin-4-ylmethyl-phenylamine (F45-000) of Example 2-2(B). Thiscompound was deprotected by a similar operation to that in the seventhstep of Example 2-13 to obtain the title compound (SH-a31).

LC/MS (Method 1): m/z (ESI, POS): 336 [M+H]⁺; retention time: 5.21minutes.

¹H-NMR [400 MHz, CDCl₃—CD₃OD (three drops)] δ 1.20 (d, J=7.0 Hz, 6H),1.52-1.64 (m, 1H), 1.84 (tt, J=7.0, 7.0 Hz, 2H), 1.96-2.06 (m, 1H), 3.21(sept., J=7.0 Hz, 1H), 3.63 (dt, J=2.6, 7.0 Hz, 2H), 4.00 (dd, J=8.8,14.0 Hz, 1H), 4.24 (dd, J=4.0, 14.0 Hz, 1H), 4.42-4.52 (m, 1H), 6.38 (s,1H), 7.32 (s, 1H).

Example 1-15 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(2-methoxy-ethyl)-2,4-dihydro-[1,2,4]triazol-3-thione(SH-a32)

The bis(methoxymethyl) protected compound of the title compound (SH-a32)was obtained in 2 steps from IM4-a by sequentially following a similarprocess to that of Example 2-12 reacting (2-methoxyethyl)thiocarbamicacid O-phenyl ester to IM4-a in place of G06-02 of Example 2-12. Thiscompound was deprotected by a similar operation to that of the seventhstep of Example 1-1 to obtain the title compound (SH-a32).

LC/MS (Method 3): m/z (ESI, POS): 310 [M+H]⁺; retention time: 4.65minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=3:1, ppm): 7.24 (1H, s), 6.37 (1s), 4.23(2H, t, J=5.86 Hz), 3.71 (3H, t, J=5.86 Hz), 3.22 (1H, m), 3.20 (3H, s),1.20 (6H, d, J=6.96 Hz).

Example 1-16 Preparation of5-(2,4-dihydroxy-5-tert-butyl-phenyl)-4-pyridin-3-yl-2,4-dihydro-[1,2,4]triazol-3-one(SH-f08)

The First Step: Preparation of 5-tert-butyl-2,4-dihydroxy-benzoic acid(F71-02)

To a tert-butanol (14.3 mL, 11.12 g, 150 mmol) suspension of2,4-dihydroxy-benzoic acid (F71-01: 2312 mg, 15.0 mmol), trifluoroaceticacid (8.0 mL, 11.84 g, 103.8 mmol) and sulfuric acid (0.43 mL, 0.79 g,8.0 mmol) were added in this order under argon atmosphere while stirringat room temperature. The reacting solution was stirred at roomtemperature for 10 minutes and then at a bath temperature of 75° C. for6 hours. Additional trifluoroacetic acid (8.0 mL, 11.84 g, 103.8 mmol)was added, and the reaction mixture was stirred at a bath temperature of80° C. for further 2.5 hours. After completing the reaction, thereaction mixture was added to ice-water (160 mL) and stirred with theaddition of hexane (20 mL). The deposited solids were collected byfiltration, washed with water and hexane to obtain the title compound(F71-02: pale pink solid, 2.13 g, 68%).

LC/MS (Method 3): m/z (ESI, POS): 209 [M−H]; retention time: 5.55minutes.

¹H-NMR (400 MHz, DMSO-D₆, TMS) ppm: 1.305 (9H, s, tert-Bu), 6.330 (1H,s, Ar—H), 7.558 (1H, s, Ar—H), 10.430 (1H, s), 11.192 (1H, bs),12.0-14.0 (b).

Preparation of5-(2,4-dihydroxy-5-tert-butyl-phenyl)-4-pyridin-3-yl-2,4-dihydro-[1,2,4]triazol-3-one(SH-f08)

The title compound was synthesized in a similar manner to Example 1-6 byusing the bis(methoxymethyl) protected compound of5-tert-butyl-2,4-bis-methoxymethoxy-benzoic acid (F71-02) in place ofIM3-a.

LC/MS (Method 3): m/z (ESI, POS): 372 [M+H]⁺; retention time: 5.78minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 13.8 (1H, s), 9.66 (1H, s), 9.44(1H, s), 7.15 (2H, d, J=9.0 Hz), 6.93 (2H, d, J=9.0 Hz), 6.87 (1H, s),6.25 (1H, s), 3.75 (3H, s), 1.18 (9H, s).

Example 2-1A Production (Method A) of4-{5-hydroxy-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol(OH-a01)

The First Step: Preparation of4-[4-(morpholin-4-yl)-phenyl]-1-[5-isopropyl-2,4-bis(methoxymethoxy)-benzoyl]semicarbazide(IM5-O-a01)

To a solution of 4-[4-(morpholin-4-yl)-phenyl]semicarbazide(IM9-O-01:70.9 mg, 0.3 mmol) and5-isopropyl-2,4-bis(methoxymethoxy)-benzoic acid (IM3-a : 85.3 mg, 0.3mmol) in dimethylformamide (4 mL), were added N-methylpyrrolidone (0.2mL), 1-hydroxybenzotriazole monohydrate (60.8 mg, 0.45 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (115.0 mg, 0.6 mmol)successively, and the mixture was stirred at room temperature overnight.The reaction mixture was mixed with ethyl acetate (30 mL) and 5% aqueoussodium hydrogencarbonate (20 mL). The aqueous layer was separated, andextracted with ethyl acetate. The combined organic layers thus obtainedwere dried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration, the solvent was distilled off under reduced pressure, andthe residue was purified by silica gel column chromatography(dichloromethane: methanol=30:1-20:1) to obtain the title compound(IM5-O-a01: colorless syrup, 104.9 mg, 70%).

LC/MS (Method 1): m/z (ESI, POS): 503 [M+H]⁺; retention time: 5.88minutes.

The Second Step: Preparation of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-ol(IM6-O-a01)

4-[4-(morpholin-4-yl)-phenyl]-1-[5-isopropyl-2,4-bis(methoxymethoxy)-benzoyl]semicarbazide(IM5-O-a01: 104.9 mg, 0.209 mmol) was mixed with 5% aqueous sodiumhydroxide solution (2 mL) and stirred 105° C. (bath temperature) for 2hours. Further, potassium hydroxide (100 mg) was added to the reactionmixture and the resulting mixture was stirred at 130° C. (bathtemperature) for 3 hours. After cooling to room temperature, thereaction mixture was neutralized with 2 N hydrochloric acid and aqueoussodium hydrogencarbonate, extracted with dichloromethane (50 mL) anddried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration, the solvent was distilled off under reduced pressure, andthe residue was purified by silica gel column chromatography(dichloromethane: methanol=20:1) to obtain the title compound(IM6-O-a01: colorless syrup, 16.3 mg, 16%).

LC/MS (Method 1): m/z (ESI, POS): 485 [M+H]⁺; retention time: 5.89minutes.

The Third Step: Preparation of4-{5-hydroxy-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol(OH-a01)

An ethanol solution (2 mL) of1,3-bis(methoxymethoxy)-4-{5-hydroxy-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropylbenzene(IM6-O-a01: 16.3 mg, 0.034 mmol) was mixed with 5 N hydrochloric acid (1mL) and reacted at room temperature for 4 hours. The solvent wasdistilled off under reduced pressure, and the residue was neutralizedwith 5% sodium hydrogencarbonate and extracted with dichloromethane (30mL). The extract was dried over anhydrous sodium sulfate. Sodium sulfatewas removed by filtration, the solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (dichloromethane: methanol=30:1-20:1) to obtain the titlecompound (OH-a01: white solid, 3.0 mg, 22%).

LC/MS (Method 1): m/z (ESI, POS): 397 [M+H]⁺; retention time: 5.10minutes.

¹H-NMR (200 MHz, CDCl₃+CD₃OD=2:1, TMS) ppm: 0.79 (6H, d, J=6.8 Hz), 2.19(1H, sept, J=6.8 Hz), 3.15-3.24 (4H, m), 3.80-3.92 (4H, m), 6.39 (1H,s), 6.54 (1H, s), 7.03 (2H, d, J=9.1 Hz), 7.21 (2H, d, J=9.0 Hz).

Example 2-1B Production (Method B) of4-{5-hydroxy-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol(OH-a01)

The First Step: Preparation of2,4-bis-benzyloxy-5-isopropyl-N-[4-(morpholin-4-yl)-phenyl]benzamide(F370-IM2)

A dichloromethane (30 mL) solution of2,4-bis-benzyloxy-5-isopropylbenzoic acid (F370-IM1, 2 g, 5.31 mmol) wasmixed with dimethylformamide (0.053 mL, 0.05 mmol) and oxalylchloride(0.61 mL, 6.38 mmol) under ice cold conditions and the mixture wasstirred at room temperature overnight. The reaction mixture wasconcentrated under reduced pressure, and the residue thus obtained wasdissolved in tetrahydrofuran (30 mL) and pyridine (10 mL). To thesolution was added 4-(4-morpholino)aniline (1.04 g, 5.84 mmol) under icecold conditions. The reaction was carried out at room temperature for 1hour. Dichloromethane (50 mL) and 5% aqueous sodium hydrogencarbonate(20 mL) were added to the reaction mixture. The aqueous layer wasseparated and extracted with dichloromethane, the combined organiclayers thus obtained were washed with saturated aqueous sodium chloridesolution (30 mL) and dried over anhydrous sodium sulfate. Sodium sulfatewas removed by filtration, and the solvent was distilled off underreduced pressure. To the residue were added ethyl acetate (5 mL),toluene (5 mL) and hexane (20 mL), and the resulting suspension wasstirred at room temperature for 1 hour. The precipitated solid wascollected by filtration to obtain the title compound (F370-IM2: whitecrystals, 2.47 g, 87%).

LC/MS (Method 3): m/z (ESI, POS): 537 [M+H]⁺; retention time: 8.61minutes.

The Second Step: Preparation of2,4-bis-benzyloxy-5-isopropyl-N-[4-(morpholin-4-yl)-phenyl]thiobenzamide(F370-IM3)

2,4-bis-benzyloxy-5-isopropyl-N-[4-(morpholin-4-yl)phenyl]benzamide(F370-IM2, 2.45 g, 4.60 mmol) was suspended in toluene (50 mL), mixedwith Lawesson's reagent (2.05 g, 5.06 mmol) and the mixture was heatedat 110° C. for 1 hour. As the reaction proceeded, the suspension changedto a yellow solution. The reaction mixture was cooled to roomtemperature, mixed with 5% aqueous sodium hydrogencarbonate (50 mL), andthe organic layer was separated. The organic layer thus obtained wasdried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration, and the solvent was distilled off under reduced pressure. Tothe residue were added toluene (20 mL) and hexane (10 mL), and theresulting suspension was stirred at room temperature for 1 hour,crystals (F370-IM3) were collected by filtration. The mother liquor wasconcentrated and purified by silica gel column chromatography (ethylacetate:hexane 2:1), to afford F370-IM3. The combined title compound(F370-IM3, about 4.1 g) was subjected to the next reaction.

LC/MS (Method 3): m/z (ESI, POS): 553 [M+H]⁺; retention time: 8.72minutes.

The Third Step: Preparation of2,4-bis-benzyloxy-5-isopropyl-N-[4-(morpholin-4-yl)-phenyl]-benzene-carbohydrazonamide(F370-IM4)

2,4-bis-benzyloxy-5-isopropyl-N-[4-(morpholin-4-yl)-phenyl]thiobenzamide(F370-IM3; crude product, 4.2 g) was suspended in ethanol (30 mL), mixedwith 80% aqueous hydrazine solution (15 mL) and heated under reflux for4 hours. At this time, it was observed that the suspension was decoloredand became a homogeneous solution. After cooling to room temperature,the reaction mixture was concentrated under reduced pressure and theresidue thus obtained (crude F370-IM4) was used for the next reactionwithout purification.

LC/MS (Method 3): m/z (ESI, POS): 551 [M+H]⁺; retention time: 4.67minutes.

The Fourth Step: Preparation of5-(2,4-bis-benzyloxy-5-isopropyl-phenyl)-4-(4-morpholin-4-yl-phenyl)-4H-[1,2,4]triazol-3-ol(F370-IM5)

2,4-bis-benzyloxy-5-isopropyl-N-[4-(morpholin-4-yl)-phenyl]-benzene-carbohydrazonamide(F370-IM4: crude product) was suspended in tetrahydrofuran (10 mL),mixed with 1,1′-carbonyldiimidazole (1.12 g, 6.9 mmol) and the mixturewas stirred at room temperature for 2 hours. The reaction mixture waspartitioned between ethyl acetate (50 mL) and 5% aqueous sodiumhydrogencarbonate (50 mL), and the organic layer was separated. Theorganic layer obtained was dried over anhydrous sodium sulfate. Sodiumsulfate was removed by filtration, and the solvent was distilled offunder reduced pressure, and the residue thus obtained was purified bysilica gel column chromatography (ethyl acetate 100%). The obtainedproduct was purified by suspending in ethyl acetate to obtain the titlecompound (F370-IM5: white crystals, 1.2 g, 45% in 3 steps).

LC/MS (Method 3): m/z (ESI, POS): 576 [M+H]⁺; retention time: 7.37minutes.

The Fifth Step: Preparation of4-{5-hydroxy-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol(OH-a01)

5-(2,4-bis-benzyloxy-5-isopropyl-phenyl)-4-(4-morpholin-4-yl-phenyl)-4H-[1,2,4]triazol-3-ol(1.2 g, 2.07 mmol) was suspended in methanol (100 mL), acetic acid (50mL) and dimethylformamide (50 mL), and mixed with palladium carbon (60mg), and catalytic reduction was carried out under a hydrogen atmosphereat 80° C. for 1 hour and then at room temperature overnight. Aftercompleting the reaction, the catalyst was removed by filtration and thesolvent was concentrated under reduced pressure. The residue was mixedwith 5% aqueous sodium hydrogencarbonate to make it neutral and thenextracted twice with chloroform. The organic layer thus obtained wasdried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration, and the solvent was distilled off under reduced pressure,and the residue thus obtained was purified by silica gel columnchromatography (dichloromethane:methanol 15:1-10:1). The obtainedproduct was purified by suspending in ethanol to obtain the titlecompound (OH-a01: white crystals, 489 mg, 59%). The analytical data ofthis compound were the same as that of the target compound obtained inExample 2-1A.

Example 2-2(A) Preparation of4-{5-hydroxy-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-6-isopropyl-benzene-1,3-diol(OH-a02)

The First Step: Preparation of4-[4-(morpholin-4-ylmethyl)-phenyl]-1-[2,4-bis-allyloxy-5-isopropyl-benzoyl]semicarbazide(IM5-O-a02-Allyl)

The title compound (IM5-O-a02-Allyl: white crystals, 232.5 mg, 90%) wasobtained by a similar process to that of the first step of Example 2-1A,using 4-[4-(morpholin-4-ylmethyl)-phenyl]semicarbazide (IM9-O-02:125 mg,0.5 mmol) in place of 4-[4-(morpholin-4-yl)-phenyl]semicarbazide and2,4-bisallyloxy-5-isopropyl-benzoic acid (IM3-a-Allyl: 156 mg, 0.5 mmol)in place of 5-isopropyl-2,4-bis-methoxy methoxy-benzoic acid.

The second Step: Preparation of5-(5-isopropyl-2,4-bis-allyloxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-ol(IM6-O-a02-Allyl)

The title compound (IM6-O-a02-Allyl: colorless syrup, 28.8 mg, 18%) wasobtained by a similar process to that of the second step of Example2-1A, using4-[4-(morpholin-4-ylmethyl)-phenyl]-1-[2,4-bis-allyloxy-5-isopropyl-benzoyl]semicarbazide(IM5-β-a02-Allyl: 165.6 mg, 0.32 mmol) in place of4-[4-(morpholin-4-yl)-phenyl]-1-[5-isopropyl-2,4-bis-methoxymethoxy-benzoyl]semicarbazideand potassium hydroxide in place of sodium hydroxide.

LC/MS (Method 3): m/z (ESI, POS): 485 [M+H]⁺; retention time: 3.96minutes.

The Third Step: Preparation of4-[5-hydroxy-4-(4-morpholin-4-ylmethyl-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(OH-a02)

A methanol solution of5-(5-isopropyl-2,4-bis-allyloxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-ol(37.9 mg, 0.077 mmol) was mixed with tetrakis(triphenylphosphine)palladium (7 mg, 0.007 mmol) and potassium carbonate (64 mg, 0.46 mmol)and reacted at 80° C. for 12 hours. The residue obtained after theconcentration was purified by silica gel column chromatography(dichloromethane:methanol 15:1-10:1) to obtain the title compound(OH-a02: white solid, 4.5 mg, 14%).

LC/MS (Method 3): m/z (ESI, POS): 411 [M+H]⁺; retention time: 1.19minutes.

¹H-NMR (200 MHz, CDCl₃:CD₃OD=2:1, TMS) ppm: 0.79 (6H, d, J=6.8 Hz),2.48-2.55 (4H, br), 2.99 (1H, sep, J=6.8 Hz), 3.58 (2H, s), 3.74 (4H, t,J=4.6 Hz), 6.37 (1H, s), 6.55 (1H, s), 7.29 (1H, d, J=8.4 Hz), 7.50 (1H,d, J=8.2 Hz).

Example 2-2(B) Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-morpholin-4-ylmethyl-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a02) monohydrochloride

The First Step: Preparation of 4-morpholin-4-ylmethyl-phenylamine(F-45-000)

While stirring under ice cold conditions, an ethyl acetate (400 mL)solution of 4-(4-nitrobenzyl)-morpholine (25.6 g, 115.1 mmol) was addedto a concentrated hydrochloric acid (115 mL) solution of stannouschloride dihydrate (97.7 g, 402.7 mmol) dropwise over 30 minutes. Afterstirring at room temperature overnight, sodium hydroxide was added tothe reaction mixture until it became basic. The reaction mixture wasextracted with ethyl acetate and washed with saturated aqueous sodiumchloride solution. After drying the extract over anhydrous sodiumsulfate, sodium sulfate was removed by filtration, and the solvent wasdistilled off under reduced pressure to obtain a yellow solid. Afterpurification by being suspended in diethyl ether, the solid obtained wasdried under reduced pressure. The filtrate, after concentrated, waspurified by silica gel column chromatography (hexane, ethyl acetate).Combined yield of the title compound, 4-morpholin-4-ylmethyl-phenylamine(F45-000, white solid) was 16.2 g (73%).

LC/MS (Method 4): m/z (ESI, POS): 193 [M+H]⁺; retention time: 1.10minutes.

The Second Step: Preparation of 4-(4-isothiocyanatobenzyl)-morpholine(F45-00)

Triethylamine (4.5 mL, 65.4 mmol) was added to a tetrahydrofuran (500mL) solution of 4-morpholin-4-ylmethyl-phenylamine (F45-000, 5.16 g,26.8 mmol). After cooling in ice cold, thiophosgene (2.45 mL, 32.1 mmol)was added. After stirring at room temperature overnight, aqueous sodiumhydroxide was added until the reaction mixture became basic. Thereaction mixture was extracted with ethyl acetate, washed with water andthen washed with saturated sodium chloride. The extract solution wasdried over anhydrous sodium sulfate, and then sodium sulfate was removedby filtration and the solvent was distilled off under reduced pressureto obtain a red syrup like substance. The syrup like substance waspurified by silica gel column chromatography (hexane, ethyl acetate) toobtain 4-(4-isothiocyanatobenzyl)-morpholine (F45-00, brown oil, 5.72 g,91%).

LC/MS (Method 3): m/z (ESI, POS): 235 [M+H]⁺; retention time: 1.84minutes.

The Third Step: Preparation of F45-01

While stirring under ice cold conditions, an ethanol solution (5 mL) of4-(4-isothiocyanatobenzyl)-morpholine (F45-00, 2.34 g, 10.0 mmol) wasadded to an ethanol solution (4 mL) of hydrazine monohydrate (1.0 g,20.0 mmol) and stirred at room temperature for 40 minutes. The solidswere collected from the suspension by filtration and washed with hexane.The solids thus obtained were dried under reduced pressure to obtainF45-01 (pale yellow solid, 2.34 g, 88%).

LC/MS (Method 3): m/z (ESI, POS): 267 [M+H]⁺; retention time: 0.94minutes.

The Fourth Step: Preparation of F45-02

At room temperature, 4-(4,6-dimethoxy-1,3,5-triadin-2-yl)-4-methylmorpholinium chloride n hydrate (DMT-MM, 2.55 g) was added to5-isopropyl-2,4-bis-methoxymethoxybenzoic acid (2.2 g, 7.74 mmol) andF45-01 (2.16 g, 8.11 mmol) in a mixed solvent of dimethylformamide (10mL) and tetrahydrofuran (5 mL) and stirred for 5 hours. Afterterminating the reaction by adding water to the reaction mixture, thesolution was neutralized by adding saturated aqueous sodiumhydrogencarbonate. The reaction mixture was extracted with ethylacetate, and the extract was washed with water and then saturated sodiumchloride solution. After drying over anhydrous sodium sulfate, sodiumsulfate was removed by filtration, and the solvent was distilled offunder reduced pressure to obtain pale yellow solids. The solids thusobtained were suspension purified (hexane, ethyl acetate), collected byfiltration and dried under reduced pressure to obtain F45-02 (whitesolid, 3.39 g, 82%).

LC/MS (Method 3): m/z (ESI, POS): 533 [M+H]⁺; retention time: 3.80minutes.

The Fifth Step: Preparation of5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(F45-03)

At room temperature, 10% aqueous potassium hydroxide (20 mL) and 5%ethanol solution of potassium hydroxide (12 mL) were added to F45-02(3.39 g, 6.36 mmol) and then the reaction mixture was heated underreflux for 11 hours. After returning to room temperature, saturatedsodium chloride solution was added and the mixture was stirred for awhile. And then the reaction mixture was extracted with chloroform, andthe extract was washed with saturated sodium chloride solution. Afterdrying the extract over anhydrous sodium sulfate, sodium sulfate wasremoved by filtration and the solvent was distilled off under reducedpressure to obtain pale yellow solids. It was purified by silica gelcolumn chromatography (NH silica, hexane, ethyl acetate, chloroform andmethanol) to obtain5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(F45-03, white solid, 2.43 g, 74%).

LC/MS (Method 3): m/z (ESI, POS): 515 [M+H]⁺; retention time: 3.56minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.11 (d, J=7.0 Hz, 6H), 2.42 (t, J=4.6 Hz,4H), 3.19 (sept., J=7.0 Hz, 1H), 3.26 (s, 3H), 3.46 (s, 3H), 3.47 (s,2H), 3.70 (t, J=4.6 Hz, 4H), 4.74 (s, 2H), 5.16 (s, 2H), 6.81 (s, 1H),7.09 (s, 1H), 7.23 (d, J=8.4 Hz, 2H), 7.35 (d, J=8.4 Hz, 2H).

The Sixth Step: Preparation of3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(F45-04)

5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(F45-03, 2.43 g, 4.72 mmol) and potassium carbonate (653 mg, 4.72 mmol)were weighed and placed in a reaction vessel, and ethanol (30 mL) andthen methyl iodide (0.29 mL, 4.72 mmol) were added. After heating at 80°C. stirring for an hour, the reaction mixture was returned to roomtemperature, and the solvent was distilled off under reduced pressure.The reaction system was mixed with water, extracted with ethyl acetate,and the extract was washed with saturated sodium chloride solution.After drying the extract over anhydrous sodium sulfate, sodium sulfatewas removed by filtration and the solvent was distilled off underreduced pressure to obtain a pale yellow foam (2.18 g) containing3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(F45-04).

LC/MS (Method 3): m/z (ESI, POS): 529 [M+H]⁺; retention time: 3.47minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.13 (d, J=7.0 Hz, 6H), 2.42 (t, J=4.6 Hz,4H), 2.73 (s, 3H), 3.19 (s, 3H), 3.21 (sept., J=7.0 Hz, 1H), 3.46 (s,3H), 3.48 (s, 2H), 3.70 (t, J=4.6 Hz, 4H), 4.70 (s, 2H), 5.15 (s, 2H),6.78 (s, 1H), 7.10 (d, J=8.4 Hz, 2H), 7.25 (s, 1H), 7.34 (d, J=8.4 Hz,2H).

The Seventh Step: Preparation of3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-5-methanesulfonyl-4H-[1,2,4]triazole(F45-05)

While stirring under ice cold conditions, 3-chloroperbenzoic acid (3.54g, 20.5 mmol) was added to a methylene chloride solution (20 mL) of thecrude product (2.15 g), obtained in the previous step, containing3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(F45-04). After stirring at room temperature for 3.5 hours, additional3-chloroperbenzoic acid (701 mg, 4.06 mmol) was added to the reactionmixture and further stirred for 1.5 hours. After cooling to ice coldtemperature, saturated aqueous sodium thiosulfate and then 10% aqueouspotassium hydrogen sulfite were added, and the mixture was stirred forsome time. Further, saturated aqueous sodium hydrogencarbonate wasadded, and the mixture was stirred and then extracted with ethylacetate. The extract was washed with saturated sodium chloride solutionand dried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration and the solvent was distilled off under reduced pressure toobtain a red brown foam (2.49 g) containing3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-5-methanesulfonyl-4H-[1,2,4]triazole(F45-05).

LC/MS (Method 3): m/z (ESI, POS): 561 [M+H]⁺; retention time: 3.63minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.11 (d, J=7.0 Hz, 6H), 2.41 (t, J=4.6 Hz,4H), 3.20 (sept., J=7.0 Hz, 1H), 3.25 (s, 3H), 3.46 (s, 3H), 3.48 (s,2H), 3.54 (s, 3H), 3.69 (t, J=4.6 Hz, 4H), 4.78 (s, 2H), 5.17 (s, 2H),6.82 (s, 1H), 7.18 (s, 1H), 7.26 (d, J=8.4 Hz, 2H), 7.36 (d, J=8.4 Hz,2H).

The Eighth Step: Preparation of5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazole-3-one(F45-06)

At room temperature aqueous sodium hydroxide (1.0 M aqueous solution, 7mL) was added to a dimethylsulfoxide solution (7 mL) of the crudeproduct (1.01 g), which was obtained in the previous step, containing3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-5-methanesulfonyl-4H-[1,2,4]triazole(F45-05). The reaction mixture was heated at 90° C. for 5.5 hours whilestirring, cooled to room temperature, extracted with ethyl acetate, andthe extract was washed twice with water and then saturated sodiumchloride solution. The extract was dried over anhydrous sodium sulfate,and then sodium sulfate was removed by filtration. The solvent wasdistilled off under reduced pressure to obtain a pale yellow foam.Reprecipitation purification was carried out (chloroform, diethylether), and the solids were washed with diethyl ether and dried underreduced pressure to obtain5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazole-3-one(F45-06, pale yellow solid, 469 mg, 53%).

LC/MS (Method 3): m/z (ESI, POS): 499 [M+H]⁺; retention time: 3.41minutes.

The Ninth Step: Preparation of5-(2,4-dihydroxy-5-isopropylphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazole-3-one(OH-a02)

5 N hydrochloric acid (2 mL) was added to a methanol solution (5 mL) of5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazole-3-one(F45-06, 469 mg, 0.94 mmol) and the mixture was stirred at roomtemperature for 3 hours. An additional 5 N hydrochloric acid (2 mL) wasadded and the stirring was continued overnight. The reaction mixture wascooled to ice cold conditions and neutralized by adding saturatedaqueous sodium hydrogencarbonate. The solution was extracted with amixed solvent of chloroform-methanol, and the extract was washed withwater and saturated sodium chloride solution. After drying the extractover anhydrous sodium sulfate, sodium sulfate was removed by filtrationand the solvent was distilled off under reduced pressure to obtain whitesolids. The solids thus obtained were dissolved in methanol (20 mL) andmethylene chloride (100 mL) and insoluble components were removed byfiltration. The mother liquor after the filtration was concentratedunder reduced pressure, and the solids obtained were purified by silicagel column chromatography (methanol:methylene chloride=1:9) to obtainthe title compound (537.7 mg).

LC/MS (Method 3): m/z (ESI, POS): 411 [M+H]⁺; retention time: 1.69minutes.

¹H-NMR (400 MHz, DMSO-d₆) δ 0.94 (d, J=7.0 Hz, 6H), 2.33 (s, 4H), 2.96(sept., J=7.0 Hz, 1H), 3.44 (s, 2H), 3.56 (t, J=4.6 Hz, 4H), 6.32 (s,1H), 6.76 (s, 1H), 7.13 (d, J=8.4 Hz, 2H), 7.30 (d, J=8.4 Hz, 2H), 9.45(bs, 1H), 9.70 (bs, 1H), 11.93 (bs, 1H).

The Tenth Step: Preparation of5-(2,4-dihydroxy-5-isopropylphenyl)-4-(4-morphlin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a02) monohydrochloride

5-(2,4-dihydroxy-5-isopropylphenyl)-4-(4-morpholin-4-ylmethylphenyl)-2,4-dihydro-[1,2,4]triazole-3-on(OH-a02, 532 mg, 1.3 mmol) and 1,4-dioxane (50 mL) were placed in a 200mL eggplant shaped flask, and 4 N hydrochloric acid/1,4-dioxane solution(0.33 mL, 1.3 mmol) was added at room temperature while stirring. Thestirring was continued at room temperature for further 2 hours. Aftercompleting the reaction, most of the 1,4-dioxane was removed underreduced pressure, and the reaction mixture was purified by beingsuspended in diethyl ether. The solids were collected by filtration anddried under reduced pressure to obtain the title compound (OH-02amonohydrochloride, 542.1 mg).

LC/MS (Method 4): m/z (ESI, POS): 411 [M−HCl+H]⁺; retention time: 3.81minutes.

¹H-NMR (400 MHz, DMSO-d₆) δ 1.01 (d, J=6.8 Hz, 6H), 3.01 (sept., J=6.8Hz, 1H), 3.08 (m, 2H), 3.19 (m, 2H), 3.72 (m, 2H), 3.94 (m, 2H), 4.32(m, 2H), 6.27 (s, 1H), 6.89 (s, 1H), 7.25 (d, J=7.7 Hz, 2H), 7.56 (d,J=7.7 Hz, 2H), 9.36 (s, 1H), 9.64 (s, 1H), 10.85 (bs, 1H), 11.97 (s,1H).

Example 2-3A and Example 2-3B

Example 2-3A Production (Method A) of5-[5-(but-2-ynyl)-2,4-dihydroxy-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-c02)

The title compound (OH-c02) was prepared from2,4-bis-allyloxy-5-(but-2-ynyl)-benzoic acid (IM07-c) via F470-IM13 in asimilar manner to Example 2-2. The MS, and NMR spectral data of thetitle compound (OH-a02) are described in Example 2-3B.

Example 2-3B Preparation (Method B) of5-[5-(but-2-ynyl)-2,4-dihydroxy-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2-4]triazol-3-one(OH-c02) The First Step: Preparation of2,4-bis-allyloxy-5-(but-2-ynyl)-N-[4-(morpholin-4-ylmethyl)-phenyl]-benzamide(F470-IM09)

2,4-bis-allyloxy-5-(but-2-ynyl)-benzoic acid (Example 1-4, IM07-c: 287mg, 1 mmol) and 4-(morpholin-4-ylmethyl)-phenylamine (IM08-02: 193 mg, 1mmol) were dissolved in dimethylformamide (4 mL) and mixed with1-hydroxybenzotriazole monohydrate (176 mg, 1.3 mmol) and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (249 mg, 1.3mmol) at 0° C., and the resulting mixture was stirred for 4 hours underice cold conditions. The reaction mixture was mixed with water (40 mL)and extracted twice with ethyl acetate (50 mL). The combined organiclayers were washed with saturated aqueous sodium chloride solution,dried over sodium sulfate and then were concentrated. The residue waspurified by silica gel column chromatography(hexane-ethylacetate=1:2-1:5) to obtain the title compound (F470-IM09:0.40 g, yield 82%).

LC/MS: m/z (ESI, POS): 461 [M+H]⁺

¹H-NMR (400 MHz, CDCl₃, TMS) ppm: 9.90 (1H, s), 8.39 (1H, s), 7.63 (2H,d, J=8.4 Hz), 7.36 (2H, d, J=8.4 Hz), 6.46 (1H, s), 6.18 (2H, m), 6.05(2H, m), 5.57-5.31 (4H, m), 4.70 (2H, m), 4.61 (2H, m), 3.80 (4H, brs),3.65 (2H, m), 2.62 (4H, brs), 4.61 (2H, m), 3.46 (2H, m), 1.85 (3H, t,J=2.6 Hz).

The Second Step: Preparation of2,4-bis-allyloxy-5-(but-2-ynyl)-N-[4-(morpholin-4-ylmethyl)-phenyl]-thiobenzamide(F470-IM10)

2,4-bis-allyloxy-5-(but-2-ynyl)-N-[4-(morpholin-4-ylmethyl)-phenyl]-benzamide(F470-IM09: 400 mg, 0.86 mmol) and Lawesson's reagent (352 mg, 0.86mmol) were dissolved in toluene (20 mL), and the mixture was heatedunder reflux for 3 hours. The reaction mixture was mixed with saturatedaqueous sodium carbonate (30 mL) and extracted twice with ethyl acetate(40 mL). The combined organic layers were washed with saturated aqueoussodium chloride solution, dried over anhydrous sodium sulfate and thenconcentrated. The residue was purified by silica gel columnchromatography (ethyl acetate) to obtain the crude title compound(F470-IM10:240 mg).

LC/MS: m/z (ESI, POS): 477 [M+H]⁺

The Third Step: Preparation of2,4-bis-allyloxy-5-(but-2-ynyl)-N-[4-(morpholin-4-yl)-phenyl]-benzene-carbohydrazonamide(F470-IM11)

Crude2,4-bis-allyloxy-5-(but-2-ynyl)-N-[4-(morpholin-4-ylmethyl)-phenyl]-thiobenzamide(F470-IM10: 240 mg, 0.86 mmol) and hydrazine•monohydrate (700 mg, 14mmol) were dissolved in ethanol (6 mL) and the mixture was heated underreflux for 1 hour and then concentrated to obtain the crude titlecompound (F470-IM11: 243 mg).

LC/MS (Method 4): m/z (ESI, POS): 475 [M+H]⁺; retention time: 3.88minutes.

The Fourth Step: Preparation of5-[2,4-bis-allyloxy-5-(but-2-ynyl)-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F470-IM12)

Crude2,4-bis-allyloxy-5-(but-2-ynyl)-N-[4-(morpholin-4-yl)-phenyl]-benzene-carbohydrazonamide(F470-IM11: 243 mg) was dissolved in anhydrous tetrahydrofuran (5 mL)and mixed with 1,1′-carbonyldiimidazole (121 mg, 0.75 mmol), and themixture was stirred for 1.5 hours. The reaction mixture was mixed withsaturated sodium carbonate solution (15 mL) and extracted twice withethyl acetate (30 mL). The organic layer was washed with saturatedsodium chloride solution, dried over anhydrous sodium sulfate and thenconcentrated to obtain the crude title compound (F470-IM12: 208 mg).

LC/MS (Method 4): m/z (ESI, POS): 501 [M+H]⁺; retention time: 4.90minutes.

The Fifth Step: Preparation of5-[5-(but-2-ynyl)-2,4-dihydroxy-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-c02)

Under a nitrogen atmosphere crude5-[2,4-bis-allyloxy-5-(but-2-ynyl)-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F470-IM12: 208 mg) was dissolved in methanol (10 mL) and mixed withpotassium carbonate (348 mg, 2.52 mmol) and tetrakis(triphenylphosphine)palladium (20 mg, 0.016 mmol), and the mixture was heated for 3 hoursunder reflux. After adding water (5 mL), the reaction mixture wasadjusted to pH 6.5 with 2M hydrochloric acid. Silica gel (2.0 g) wasadded to this mixture and the mixture was concentrated. The residue waspurified by silica gel column chromatography(chloroform-methanol=30:1-10:1) to obtain the title compound (OH-c02: 38mg, yield 10.4%, in 4 steps).

LC/MS (Method 3): m/z (ESI, POS): 421 [M+H]⁺; retention time: 1.15minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=4:1, TMS) ppm: 7.46 (2H, d, J=8.1), 7.26(2H, d, J=8.1), 6.83 (1H, s), 6.35 (1H, s), 3.75 (4H, brs), 3.40 (2H,s), 3.16 (2H, m), 2.58 (4H, brs), 1.74 (3H, t, J=2.6 Hz).

Example 2-4 Preparation of5-[2,4-dihydroxy-5-(prop-2-ynyl)-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-e02)

The First Step: Preparation of F59-02

Under a nitrogen atmosphere, 2,4-dihydroxybenzaldehyde (F59-01, 3.0 g,21.72 mmol) was suspended in anhydrous dichloromethane (300 mL) in a 1 Lflask. Benzyltrimethylammonium tribromide (10.0 g, 25.64 mmol) was addedto this mixture and the mixture was stirred at room temperature for 2.5hours. The reaction mixture was mixed with water (200 mL) and chloroform(100 mL). The organic layer was separated and washed with saturatedaqueous sodium chloride solution, dried with sodium sulfate and, afterfiltration, concentrated under reduced pressure to obtain the crudetitle compound (F59-02: 5250 mg).

The Second Step: Preparation of F59-03

Crude F59-02 (5250 mg) was dissolved in anhydrous dimethylformamide (50mL) and under ice cold conditions mixed with potassium carbonate (7.56g, 54.68 mmol) and benzylbromide (5.42 mL, 45.57 mmol), and the mixturewas stirred overnight. The reaction mixture was mixed with water (500mL) and extracted twice with ethyl acetate (300 mL). The combinedorganic layers were washed with saturated aqueous sodium chloridesolution, then dried with sodium sulfate and concentrated. The residuewas purified by silica gel column chromatography (hexane:ethylacetate=4:1) to obtain the title compound (F59-03: 6.2 g, yield 72.0% in2 steps).

LC/MS (Method 3): m/z (ESI, POS): 399 [M+H]⁺; retention time: 7.62minutes.

¹H-NMR (400 MHz, CDCl₃, ppm): 10.30 (1H, s), 8.04 (1H, s), 7.41-7.37(12H, m), 6.53 (1H, s), 5.16 (2H, s), 5.11 (2H, s)

The Third Step: Preparation of F59-04

Trimethylsilylacetylene (1.04 mL, 7.5 mmol) and anhydroustetrahydrofuran (15 mL) were placed in a 100 mL three-neck flask under aatmosphere of nitrogen. n-Butyllithium (4.72 mL, 1.59M/hexane) wereadded to this mixture dropwise over 30 minutes at −78° C. A solution ofF59-03 (1987 mg, 5 mmol) in tetrahydrofuran (30 mL) was added to thismixture dropwise over 20 minutes at −78° C. After being stirred for 3hours under ice cold conditions, the reaction mixture was mixed withsaturated aqueous ammonium chloride (30 mL) and water, and extractedtwice with ethyl acetate (100 mL). The combined organic layers werewashed with saturated aqueous sodium hydrogencarbonate and saturatedaqueous sodium chloride solution, and then dried with sodium sulfate.The mixture was filtered and the filtrate was concentrated under reducedpressure to obtain the crude title compound (F59-04, 2770 mg).

LC/MS (Method 7): m/z (ESI, POS): 477 [M−H₂O]⁺; retention time: 7.36minutes.

The Fourth Step: Preparation of F59-05

Under a nitrogen atmosphere, crude F59-04 (2770 mg) dissolved inanhydrous acetonitrile (4 mL) was placed in a 200 mL two-neck flask. Tothe solution were added triethylsilane (0.878 mL, 5.5 mmol) andborontrifluoride diethylether (0.697 mL, 5.5 mmol) and this mixture wasstirred for 1 hour under ice cold conditions. The reaction mixture wasmixed with potassium carbonate (2000 mg) and water (150 mL) andextracted twice with ethyl acetate (150 mL). The organic layer waswashed with saturated aqueous sodium chloride solution, dried withsodium sulfate and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (hexane:ethyl acetate=5:1)to obtain the title compound (F59-05: 2135 mg, yield 89% in 2 steps).

The Fifth Step: Preparation of F59-06

Under a nitrogen atmosphere, F59-05 (2135 mg, 4.45 mmol) dissolved inanhydrous tetrahydrofuran (20 mL) was placed in a 100 mL two-neck flask.n-Butyl lithium (3.08 mL, 1.59M/hexane, 4.90 mmol) was added to thismixture dropwise over 10 minutes at −78° C. And then the mixture wasstirred for 15 minutes. A large excess of solid carbon dioxide wasquickly added to the reaction mixture and the mixture was stirred atroom temperature for 1 hour. The reaction mixture was mixed with 10%aqueous potassium hydrogen sulfate (50 mL), extracted twice with ethylacetate (100 mL). After washing with saturated sodium chloride solution,the organic layer was dried with sodium sulfate, filtered andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane:ethyl acetate=4:1-2:1) to obtain thetitle compound (F59-06: 370 mg, yield 18.7%).

LC/MS (Method 3): m/z (ESI, POS): 445 [M+H]⁺; retention time: 8.26minutes.

¹H-NMR (400 MHz, CDCl₃, ppm): 10.52 (1H, brs), 8.28 (1H, s), 7.40 (10H,s), 6.56 (1H, s), 5.19 (2H, s), 5.12 (2H, s), 3.58 (2H, s), 0.19 (9H, s)

The Sixth Step: Preparation of F59-07

A solution of F59-06 (370 mg, 0.83 mmol) and F59-11 (192 mg, 1.0 mmol)in dimethylformamide (5 mL) was placed in a 100 mL flask.1-Hydroxybenzotriazole monohydrate (146 mg, 1.08 mmol) and1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide hydrochloride (207 mg,1.08 mmol) were added to the solution under ice cold conditions and themixture was stirred for 20 hours. The reaction mixture was mixed withwater (30 mL) and saturated aqueous sodium hydrogencarbonate (20 mL) andextracted twice with ethyl acetate (50 mL). The organic layer was washedwith saturated aqueous sodium chloride solution, dried with sodiumsulfate and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (chloroform:methanol=30:1)to obtain the title compound (F59-07: 447 mg, yield 87.0%).

LC/MS (Method 5): m/z (ESI, POS): 619 [M+H]⁺; retention time: 8.26minutes.

The Seventh Step: Preparation of F59-08

A solution of F-59-07 (477 mg, 0.72 mmol) and Lawesson's reagent (381mg, 0.94 mmol) in toluene (15 mL) was placed in a 100 mL flask, and themixture was heated for 8 hours under reflux. The reaction mixture wasmixed with saturated aqueous sodium hydrogencarbonate solution (30 mL)and extracted twice with ethyl acetate (30 mL). The organic layer waswashed with saturated sodium chloride solution, dried with sodiumsulfate and then concentrated. The residue was purified by silica gelcolumn chromatography (chloroform:methanol=40:1) to obtain the titlecompound (F59-08: 339 mg, yield 74.1%).

LC/MS (Method 5): m/z (ESI, POS): 635 [M+H]⁺; retention time: 4.52minutes.

The Eighth Step: Preparation of F59-09

F59-08 (339 mg, 0.53 mmol) and hydrazine•monohydrate (0.7 mL) weredissolved in ethanol (6 mL) and heated for 1 hour under reflux, and thenthe mixture was concentrated to obtain the crude title compound (F59-09,345 mg).

LC/MS (Method 3): m/z (ESI, POS): 633 [M+H]⁺; retention time: 3.79minutes.

The Ninth Step: Preparation of F59-10

Crude F59-09 (345 mg) was dissolved in anhydrous tetrahydrofuran (6 mL),mixed with 1-1′ carbonyldiimidazole (130 mg, 0.8 mmol) and stirred for 5hours. The reaction mixture was mixed with saturated aqueous sodiumcarbonate solution (30 mL) and extracted twice with ethyl acetate (30mL). The organic layer was washed with saturated aqueous sodium chloridesolution, dried with sodium sulfate and then concentrated. The residuewas purified by silica gel column chromatography (ethylacetate:acetone=8:1) to obtain the title compound (F59-10: 101 mg, yield21.3% in 3 steps).

LC/MS (Method 3): m/z (ESI, POS): 659 [M+H]⁺; retention time: 4.99minutes.

¹H-NMR (400 MHz, CDCl₃, ppm): 10.33 (1H, s), 7.44 (1H, s), 7.30-7.08(10H, m), 6.93 (2H, d, J=8.24 Hz), 6.84 (2H, m), 6.13 (1H, s), 4.80 (2H,s), 4.47 (2H, s), 3.59 (4H, s), 3.49 (2H, s), 3.37 (2H, s), 2.37 (4H,s), 0.06 (9H, s)

The Tenth Step: Preparation of5-[2,4-dihydroxy-5-(prop-2-ynyl)-phenyl]-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-e02)

F59-10 (101 mg, 0.15 mmol) was dissolved in anhydrous tetrahydrofuran (5mL) in a 50 mL flask, mixed with tetrabutylammonium fluoride (0.16 mL,1.0M/tetrahydrofuran) and stirred for 1 hour. The reaction mixture wasmixed with saturated aqueous sodium hydrogencarbonate (20 mL) andextracted twice with ethyl acetate (20 mL). The organic layer was washedwith saturated aqueous sodium chloride solution, dried with sodiumsulfate, filtered and then concentrated. The residue was dissolved inanhydrous dichloromethane (3 mL), mixed with boron trichloride (3 mL,1.0M/dichloromethane) at −20° C. and then stirred for 2 hours under icecold conditions. After adding methanol (5 mL) to the reaction mixture,solid sodium hydrogencarbonate was added so that the pH of the solutionwas adjusted to 7.0 by pH-test paper. The reaction mixture was filteredand the filter cake was washing sufficiently with chloroform:methanol(3:1). The filtrate and washing were combined and concentrated. Theresidue was purified by silica gel column chromatography(chloroform:methanol=10:1-5:1) to obtain the title compound (OH-e02:21.5 mg, yield 35.2%).

LC/MS (Method 4): m/z (ESI, POS): 407 [M+H]⁺; retention time: 3.58minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=3:1, ppm): 7.47 (2H, d, J=8.06 Hz), 7.27(2H, d, J=8.06 Hz), 6.88 (1H, s), 6.38 (1H, s), 3.75 (4H, brs), 3.59(2H, s), 3.22 (2H, m), 2.54 (4H, brs), 1.94 (1H, s)

Example 2-5 Preparation of5-(2,4-dihydroxy-5-isopropylphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a13) dihydrochloride

The First Step: Preparation of 1-methyl-4-(4-nitrobenzyl)-piperazine(F652-01)

Monomethylpiperazine (15 mL) and tetrahydrofuran (60 mL) were placed ina 200 mL eggplant shaped flask, and a solution of 4-nitrobenzylchloride(8.58 g, 50 mmol) in tetrahydrofuran was added dropwise to the mixtureat room temperature while stirring. After finishing the instillation,the mixture was stirred at room temperature for 24 hours. The reactionmixture was mixed with distilled water, and the precipitated solids werecollected by filtration and dried under reduced pressure to obtain thetitle compound (5.9 g, 50%).

LC/MS (Method 3): m/z (ESI, POS): 236 [M+H]⁺; retention time: 1.28minutes.

The Second Step: Preparation of4-(4-methylpiperazin-1-ylmethyl)phenylamine (F652-02)

1-methyl-4-(4-nitrobenzyl)-piperazine (F652-01, 4.67 g, 19.9 mmol),methanol (100 mL), zinc powder (6.5 g, 99.3 mmol) and ammonium chloride(4.3 g, 79.5 mmol) were placed in a reaction vessel and heated for 2hours under reflux. After returning to room temperature, the reactionmixture was filtered through Celite. The solvent of the filtrate wasdistilled off under reduced pressure to obtain a solid. Diethyl etherwas added to the solid, and insoluble components were removed byfiltration. 4-(4-methylpiperazin-1-ylmethyl)phenylamine (F652-02, whitesolid, 3.16 g, 77%) was obtained by removing the solvent of the filtrateby distillation.

LC/MS (Method 6): m/z (ESI, POS): 206 [M+H]⁺; retention time: 1.15minutes.

¹H-NMR (400 MHz, DMSO-d₆) δ 2.12 (s, 3H), 2.29 (bs, 8H), 3.23 (s, 2H),4.93 (s, 2H), 6.49 (d, J=8.4 Hz, 2H), 6.89 (d, J=8.4 Hz, 2H).

The Third Step: Preparation of1-(4-isothiocyanatobenzyl)-4-methylpiperazine (F652-03)

Triethylamine (5.1 mL, 36.6 mmol) was added to a solution of4-(4-methylpiperazin-1-ylmethyl)phenylamine (F652-02, 3.14 g, 15.3 mmol)in tetrahydrofuran (250 mL), and after cooling by ice, thiophosgene(1.11 mL, 14.6 mmol) was added. After stirring at room temperatureovernight, aqueous sodium hydrogencarbonate was added to the mixture.The reaction mixture was extracted with ethyl acetate, and the extractwas washed with saturated sodium chloride solution, dried over anhydroussodium sulfate, filtered to remove sodium sulfate, and the solvent wasdistilled off under reduced pressure to obtain brown oil. The brown oilwas purified by silica gel column chromatography (chloroform, methanol)to obtain 1-(4-isothiocyanatobenzyl)-4-methylpiperazine (F652-03,brownish oil, 2.64 g, 70%).

LC/MS (Method 3): m/z (ESI, POS): 248 [M+H]⁺; retention time: 2.87minutes.

¹H-NMR (400 MHz, CDCl₃) δ 2.29 (s, 3H), 2.45 (bs, 8H), 3.48 (s, 2H),7.17 (d, J=8.4 Hz, 2H), 7.31 (d, J=8.4 Hz, 2H).

The Fourth Step: Preparation of F652-04

An ethanol solution (2 mL) of hydrazine monohydrate (1.07 g, 21.3 mmol)was added to an ethanol solution (15 mL) of1-(4-isothiocyanatobenzyl)-4-methylpiperazine (F652-03, 2.64 g, 10.7mmol) while stirring under ice cold conditions, and the mixture wasstirred at room temperature for 1 hour. The precipitated solids werecollected by filtration and washed with ethanol and hexane. The solidsthus obtained were dried under reduced pressure to obtain F652-04 (paleyellow solid, 2.69 g, 90%).

LC/MS (Method 4): m/z (ESI, POS): 280 [M+H]⁺; retention time: 1.32minutes.

The Fifth Step: Preparation of F652-054-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride nhydrate (DMT-MM, 3.22 g) was added at room temperature to adimethylformamide (25 mL) solution of5-isopropyl-2,4-bis-methoxymethoxybenzoic acid (2.54 g, 8.92 mmol) andF652-04 (2.62 g, 9.37 mmol) obtained in the fourth step, and the mixturewas stirred for 4 hours. After stopping the reaction by adding water,the resulting mixture was neutralized by adding saturated aqueous sodiumhydrogencarbonate. The resulting mixture was extracted with ethylacetate, and the extract was washed with water and then with saturatedsodium chloride solution. After drying over anhydrous sodium, theextract was filtered to remove sodium sulfate, and then the solvent wasdistilled off under reduced pressure to obtain pale yellow solids. Tothe solids thus obtained was added diethylether, and the resultingsuspension was stirred at room temperature. The precipitated solid wascollected by filtration, and dried under reduced pressure to obtainF652-05 (pale yellow solid, 4.02 g, 83%).

LC/MS (Method 3): m/z (ESI, POS): 546 [M+H]⁺; retention time: 3.70minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.06 (d, J=6.8 Hz, 6H), 2.32 (s, 3H), 2.49(bs, 8H), 3.16 (sept., J=6.8 Hz, 1H), 3.48 (s, 2H), 3.50 (s, 3H), 3.59(s, 3H), 5.26 (s, 2H), 5.47 (s, 2H), 6.99 (s, 1H), 7.33 (d, J=8.3 Hz,2H), 7.40 (d, J=8.4 Hz, 2H), 7.87 (s, 1H).

The Sixth Step: Preparation of5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(F652-06)

10% aqueous potassium hydroxide (15 mL) and 5% potassiumhydroxide-ethanol solution (5 mL) were added to F652-05 (4.02 g, 7.38mmol) obtained in the fifth step at room temperature. After heating themixture for 3 hours under reflux, the solvent was distilled off underreduced pressure. Saturated aqueous sodium chloride solution was addedand stirring was continued for a while, and then the reaction mixturewas extracted with chloroform. The combined chloroform extracts werewashed with saturated sodium chloride solution and dried over anhydroussodium sulfate. Filtration and evaporation gave yellow solid, which waspurified by silica gel column chromatography (chloroform, methanol) toobtain5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(F652-06, a pale yellow solid, 3.19 g, 82%).

LC/MS (Method 3): m/z (ESI, POS): 528 [M+H]⁺; retention time: 3.54minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.11 (d, J=7.0 Hz, 6H), 2.33 (s, 3H), 2.50(bs, 8H), 3.20 (sept., J=7.0 Hz, 1H), 3.26 (s, 3H), 3.46 (s, 3H), 3.49(s, 2H), 4.73 (s, 2H), 5.16 (s, 2H), 6.80 (s, 1H), 7.09 (s, 1H), 7.23(d, J=8.4 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H).

The Seventh Step:3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-5-methylsulfanyl-[1,2,4]triazole(F652-07)

5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(F652-06, 3.17 g, 6.01 mmol) and potassium carbonate (829 mg, 6.00 mmol)were weighed and placed in a reaction vessel. After adding ethanol (30mL), methyl iodide (0.374 mL, 6.01 mmol) was added to the mixture. Afterheating for 1 hour under reflux, the temperature was returned to roomtemperature, and the solvent was distilled off under reduced pressure.To the reaction mixture was added water, and the mixture was extractedwith ethyl acetate. The extract was washed with saturated sodiumchloride solution, dried over anhydrous sodium sulfate and then filteredto remove sodium sulfate, and the solvent was distilled off underreduced pressure to obtain pale yellow foam. Diethyl ether was added tothe foam and the insoluble material was removed by filtration. Afterwashing with water, the filtrate was washed with saturated sodiumchloride solution. The extract was dried with sodium sulfate, and sodiumsulfate was removed by filtration. The solvent was distilled off underreduced pressure to obtain3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-5-methylsulfanyl-[1,2,4]triazole(F652-07, pale yellow foam, 2.05 g, 63%).

LC/MS (Method 3): m/z (ESI, POS): 542 [M+H]⁺; retention time: 3.68minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.14 (d, J=7.0 Hz, 6H), 2.29 (s, 3H), 2.45(bs, 8H), 2.73 (s, 3H), 3.19 (s, 3H), 3.21 (sept., J=7.0 Hz, 1H), 3.46(s, 3H), 3.49 (s, 2H), 4.69 (s, 2H), 5.15 (s, 2H), 6.78 (s, 1H), 7.09(d, J=8.3 Hz, 2H), 7.26 (s, 1H), 7.33 (d, J=8.3 Hz, 2H).

The Eighth Step:3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-5-methanesulfonyl-[1,2,4]triazole(F652-08)

3-chloroperbenzoic acid (3.91 g, 22.7 mmol) was added to a methylenechloride (9 mL) solution of3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-5-methylsulfanyl-[1,2,4]triazole(F652-07, 2.05 g, 3.78 mmol). After stirring at room temperature for 9.5hours, additional 3-chloroperbenzoic acid (717 mg, 4.15 mmol) was addedand stirring was continued overnight. After cooling to ice cold,saturated aqueous sodium thiosulfate and then 10% aqueous potassiumbisulfite were added, and stirring was continued for a while. Further,saturated aqueous sodium hydrogencarbonate was added and the mixture wasstirred, and then extracted with chloroform. The extract was washedsequentially with saturated aqueous sodium hydrogencarbonate, water andsaturated sodium chloride solution, and dried over anhydrous sodiumsulfate. Filtration and evaporation gave pale brown foam. After addingdiethyl ether, the insoluble solids were removed by filtration, and thesolvent in the filtrate was distilled off under reduced pressure. Thesolids thus obtained were purified by silica gel column chromatography(chloroform, methanol) to obtain3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-5-methanesulfonyl-[1,2,4]triazole(F652-08, pale yellow foam, 1.08 g, 50%).

LC/MS (Method 3): m/z (ESI, POS): 574 [M+H]⁺; retention time: 3.69minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.12 (d, J=6.8 Hz, 6H), 2.29 (s, 3H), 2.44(bs, 8H), 3.20 (sept., J=6.8 Hz, 1H), 3.25 (s, 3H), 3.47 (s, 3H), 3.48(s, 2H), 3.54 (s, 3H), 4.77 (s, 2H), 5.17 (s, 2H), 6.82 (s, 1H), 7.18(s, 1H), 7.25 (d, J=8.5 Hz, 2H), 7.35 (d, J=8.5 Hz, 2H).

The Ninth Step:5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F652-09)

At room temperature, aqueous sodium hydroxide (1.0 M aqueous solution, 5mL) was added to a dimethylsulfoxide (5 mL) solution of3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-5-methanesulfonyl-[1,2,4]triazole(F652-08, 1.07 g, 1.86 mmol), and the mixture was heated for 3 hoursunder reflux. An additional aqueous sodium hydroxide (1.0 M aqueoussolution, 5 mL) was added, and the mixture was heated under flux forfurther 1 hour. After returned to room temperature, the reaction mixturewas extracted with ethyl acetate and chloroform. The combined extractswere washed twice with water and then with saturated sodium chloridesolution, and dried over anhydrous sodium sulfate, and sodium sulfatewas filtered off, and the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(NH silica, chloroform, methanol) to obtain5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F652-09, pale yellow foam, 900 mg, 95%)

LC/MS (Method 3): m/z (ESI, POS): 512 [M+H]⁺; retention time: 3.47minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.16 (d, J=6.9 Hz, 6H), 2.28 (s, 3H), 2.45(br, 8H), 3.17 (s, 3H), 3.23 (sept., J=6.9 Hz, 1H), 3.44 (s, 2H), 3.48(s, 3H), 4.63 (s, 2H), 5.17 (s, 2H), 6.81 (s, 1H), 7.13 (d, J=8.4 Hz,2H), 7.21 (s, 1H), 7.29 (d, J=8.4 Hz, 2H), 9.92 (bs, 1H).

The Tenth Step:5-(2,4-dihydroxy-5-isopropylphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-onedihydrochloride (F652-10)

At room temperature, 4 N hydrochloric acid/1,4-dioxane solution (5 mL)was added to a methanol (5 mL) solution of5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one (F652-09,644 mg, 1.26 mmol). After stirring at 40° C. for 1 hour, the solvent wasdistilled off under reduced pressure. Methanol was added to the crudeproduct thus obtained, and the resulting suspension was stirred at roomtemperature. The precipitated solid was collected by filtration toobtain5-(2,4-dihydroxy-5-isopropylphenyl)-4-[4-(4-methylpiperazin-1-ylmethyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a13) dihydrochloride (white solids).

LC/MS (Method 4): m/z (ESI, POS): 424 [M−2HCl+H]⁺; retention time: 3.81minutes.

FAB-MS: m/z (POS): 424 [M−2HCl+H]⁺: melting point: 276-277° (dec.)

¹H-NMR (400 MHz, DMSO-d₆) δ 1.02 (d, J=7.0 Hz, 6H), 2.80 (bs, 3H), 3.01(sept., J=7.0 Hz, 1H), 3.38 (br, 4H), 3.81 (br, 4H), 4.24 (br, 2H), 6.30(s, 1H), 6.90 (s, 1H), 7.22 (d, J=8.4 Hz, 2H), 7.57 (bs, 2H), 9.34 (bs,1H), 9.65 (bs, 1H), 11.97 (s, 1H).

Example 2-6 Preparation of5-(2,4-dihydroxy-5-isopropylphenyl)-4-[4-(4-(morpholin-4-carbonyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a14)

The First Step: Preparation of F61-02

At room temperature, 1-hydroxybenzotriazole n hydrate (HOBt, 496 mg) and1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EPCI, 938mg, 4.89 mmol) were added sequentially to a dimethylformamide (5 mL)solution of 2,4-bisallyloxy-5-isopropylbenzoic acid (676 mg, 2.45 mmol)and F61-01(646 mg, 2.45 mmol), and the mixture was stirred for 3.5hours. After adding water, the reaction mixture was extracted with ethylacetate. The extract as washed with saturated aqueous ammonium chloridesolution, saturated aqueous sodium hydrogencarbonate and then saturatedsodium chloride solution. The extract was dried over anhydrous sodiumsulfate, and then sodium sulfate was removed by filtration and thesolvent was distilled off under reduced pressure to obtain a pale yellowsyrup, which was purified by silica gel column chromatography (hexane,ethyl acetate, chloroform, methanol) to obtain F61-02 (pale yellow foam,1.14 g, 89%).

LC/MS (Method 3): m/z (ESI, POS): 527 [M+H]⁺; retention time: 6.13minutes.

The Second Step: Preparation of4-[3-(2,4-bis-allyloxy-5-isopropylphenyl)-5-oxo-1,5-dihydro-[1,2,4]triazol-4-yl]-benzoicacid (F61-03)

Potassium hydroxide (999 mg, 17.8 mmol) was added to an aqueous solution(10 mL) of F61-02 (1.14 g, 2.17 mmol) that was obtained in the firststep, and the mixture was heated for 14 hours under reflux. Aftercooling to room temperature, 1 N hydrochloric acid was added until thesolution became acidic (pH4-5). The reaction mixture was extracted withethyl acetate, and the extract was washed with saturated sodium chloridesolution. The extract was dried over anhydrous sodium sulfate, and thensodium sulfate was removed by filtration and the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (hexane, ethyl acetate, chloroform, methanol) andthe solids thus obtained were washed with diethyl ether to obtain4-[3-(2,4-bis-allyloxy-5-isopropylphenyl)-5-oxo-1,5-dihydro-[1,2,4]triazol-4-yl]-benzoicacid (F61-03, white solids, 67.9 mg, 7%).

LC/MS (Method 3): m/z (ESI, POS): 436 [M+H]⁺; retention time: 5.95minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.21 (d, J=7.0 Hz, 6H), 3.27 (sept., J=7.0 Hz,1H), 4.01 (d, J=5.3 Hz, 2H), 4.49 (d, J=4.9 Hz, 2H), 5.05 (dd, J=1.3,17.3 Hz, 1H), 5.11 (dd, J=1.3, 10.6 Hz, 1H), 5.29 (dd, J=1.5, 10.6 Hz,1H), 5.40 (dd, J=1.5, 17.3 Hz, 1H), 5.50-5.62 (m, 1H), 5.98-6.09 (m,1H), 6.22 (s, 1H), 7.23 (d, J=8.7 Hz, 2H), 7.31 (s, 1H), 8.00 (d, J=8.7Hz, 2H).

The Third Step: Preparation of5-(2,4-bis-allyloxy-5-isopropylphenyl)-4-[4-(morpholin-4-carbonyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F61-04)

At room temperature,4-[3-(2,4-bis-allyloxy-5-isopropylphenyl)-5-oxo-1,5-dihydro-[1,2,4]triazol-4-yl]-benzoicacid (F61-03, 30 mg, 68.9 μmol),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EPCI,14.6 mg, 76.2 μmol) and 1-hydroxybenzotriazole n hydrate (HOBt, 10.3 mg)were weighed and placed in a reaction vessel, and morpholine (6.4 μL,73.5 μmol) and tetrahydrofuran (0.5 mL) were added. Triethylamine (10.6μL, 76.1 μmol) was added and the mixture was stirred for 2.5 hours.After adding water to the reaction mixture, the solvent was distilledoff under reduced pressure. The solution was extracted with ethylacetate, and the extract was washed with saturated aqueous ammoniumchloride, saturated aqueous sodium hydrogencarbonate and then saturatedsodium chloride solution. After drying over anhydrous sodium, theextract was filtered to remove sodium sulfate, and the solvent wasdistilled off under reduced pressure to obtain a brown oil, which waspurified by silica gel column chromatography (chloroform, methanol) toobtain a crude product (34.2 mg) containing5-(2,4-bis-allyloxy-5-isopropylphenyl)-4-[4-(morpholin-4-carbonyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F61-04).

LC/MS (Method 3): m/z (ESI, POS): 505 [M+H]⁺; retention time: 5.89minutes.

The Fourth Step: Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[4-(morpholin-4-carbonyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a14)

Under argon atmosphere while stirring at −20° C., trichloroborane (1.0Mmethylene chloride solution, 0.2 mL) was added to a methylene chloride(0.2 mL) solution of the crude product (10.2 mg) containing5-(2,4-bis-allyloxy-5-isopropylphenyl)-4-[4-(morpholin-4-carbonyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F61-04) obtained in the previous step, and the stirring was continuedfor 2 hours. After this, the mixture was stirred at room temperatureovernight. After adding methanol, the reaction mixture was neutralizedby adding saturated aqueous sodium hydrogencarbonate. The solution wasextracted with chloroform and then the extract was washed with saturatedsodium chloride solution. After drying over anhydrous sodium, theextract was filtered to remove sodium sulfate, and the solvent wasdistilled off under reduced pressure. The residue was purified crudelyby thin layer chromatography (chloroform, methanol) and then purified byHPLC fractionation to obtain5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[4-(morpholin-4-carbonyl)phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a14, 0.9 mg, 10%).

LC/MS (Method 3): m/z (ESI, POS): 425 [M+H]⁺; retention time: 4.11minutes.

¹H-NMR [400 MHz, CDCl₃—CD₃OD (four drops)] δ 0.83 (d, J=6.8 Hz, 6H),3.00 (sept., J=6.8 Hz, 1H), 3.50 (br, 2H), 3.64 (br, 2H), 3.79 (br, 4H),6.53 (s, 1H), 6.56 (s, 1H), 7.38 (d, J=8.4 Hz, 2H), 7.53 (d, J=8.4 Hz,2H).

Example 2-7 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a08)

The First Step: Preparation of3-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(F60-01)

The title compound (F60-01: 2.87 g, 85.3%) was obtained by using5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-methoxy-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(F53-04: 3.26 g, 7.3 mmol) in place of 5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione,and by processing in the similar manner to the first step of Example3-1. LC/MS (Method 3): m/z (ESI, POS): 460 [M+H]⁺; retention time: 6.54minutes.

The Second Step: Preparation of3-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-5-methanesulfonyl-4-(4-methoxy-phenyl)-4H-[1,2,4]triazole(F60-02)

3-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(F60-01: 2.87 g, 6.3 mmol) and methylene chloride (60 mL) were placed ina 200 mL eggplant shaped flask, cooled to 0° C., and methylene chloridesolution of metachloroperbenzoic acid (3.77 g, 21.9 mmol) was graduallyadded in 4 portions, and the mixture was stirred for 9.5 hours. Aftercompleting the reaction, 10% aqueous potassium sulfite (100 mL) wasadded and stirring was continued for 10 minutes. After that, the organiclayer was extracted, washed twice with 1 N sodium hydroxide (50 mL),dried with sodium sulfate and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography (hexane/ethylacetate) to obtain the title compound (F60-02: 2.67 g, 87%)

¹H-NMR (400 MHz, CDCl₃, TMS) ppm: 7.22 (2H, d, J=9.0 Hz), 7.20 (1H, s),6.86 (2H, d, J=9.0 Hz), 6.83 (1H, s), 5.17 (2H, s), 4.81 (2H, s), 3.80(3H, s), 3.53 (3H, s), 3.47 (3H, s), 3.27 (3H, s), 3.22 (1H, sept, J=7.0Hz), 1.15 (6H, d, J=7.0 Hz)

LC/MS (Method 3): m/z (ESI, POS): 492 [M+H]⁺; retention time: 6.36minutes.

The Third Step: Preparation of 5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F60-03)

3-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-5-methanesulfonyl-4-(4-methoxy-phenyl)-4H-[1,2,4]triazole(F60-02: 2.5 g, 5.1 mmol), dimethylsulfoxide (50 mL) and 5% aqueoussodium hydroxide (10 mL) were placed in a 200 mL eggplant shaped flaskand stirred at 120° C. for 2 hours. After completing the reaction, thereaction mixture was cooled to 0° C., neutralized with saturated aqueousammonium chloride, and deposited solids were collected by filtration.The solids thus obtained were suspension purified by hexane to obtainthe title compound (F60-03: 2.0 g, 93%).

LC/MS (Method 3): m/z (ESI, POS): 430 [M+H]⁺; retention time: 5.89minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 12.0 (1H, brs), 7.18 (1H, s), 7.07(2H, d, J=9.0 Hz), 6.90 (2H, d, J=9.0 Hz), 6.75 (2H, s) 5.20 (2H, s),4.84 (3H, s), 3.34 (3H, s), 3.15 (1H, sept, J=6.8 Hz), 3.14 (3H, s),1.11 (6H, d, J=6.8 Hz)

The Fourth Step: Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a08)

5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one(F60-03: 1.96 g, 4.6 mmol), ethanol (75 mL) and 6 N hydrochloric acid(75 mL) were placed in a 200 mL eggplant shaped flask and stirred atroom temperature for 6 hours. After completing the reaction, distilledwater (400 mL) was added, and deposited solids were dried under reducedpressure to obtain the title compound (OH-a08: 1.4 g, 91%).

LC/MS (Method 3): m/z (ESI, POS): 342 [M+H]⁺; retention time: 4.94minutes.

¹H-NMR (400 MHz, CD₃OD, TMS) ppm: 7.19 (2H, d, J=9.1 Hz), 7.00 (2H, d,J=9.1 Hz), 6.70 (1H, s), 6.27 (1H, s), 3.02 (1H, sept, J=6.8 Hz), 0.91(6H, d, J=6.8 Hz)

IR(KBr): 1708, 1627, 1514, 1394, 1302, 1253, 1174, 604.

Melting point: 272° C. (decomposition)

Example 2-8 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(3-methoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a09)

5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[3-methoxy-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thionewas obtained using 3-methoxy-phenyl isothiocyanate in place of F53-01 ofExample 1-6 by the similar process to that of Example 1-6 in 3 steps.This compound was led to the title compound (OH-a09) by the similarprocess to that of Example 2-7 in 4 steps.

LC/MS (Method 1): m/z (ESI, POS): 342 [M+H]⁺; retention time: 5.39minutes.

Example 2-9 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(3,4-dimethoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a10)

5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(3,4-dimethoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-thionewas obtained using 3,4-dimethoxy-phenyl isothiocyanate in place ofF53-01 of Example 1-6 by the similar process to that of Example 1-6 in 3steps. This compound was led to the title compound (OH-a10) by thesimilar process to that of Example 2-7 in 4 steps.

LC/MS (Method 1): m/z (ESI, POS): 373 [M+H]⁺; retention time: 4.98minutes.

Example 2-10 Preparation of5-(2,4-dihydroxy-5-isopropylphenyl)-4-(4-hydroxyphenyl)-4H-[1,2,4]triazol-3-one(OH-a11)

Crystals (3.30 g, 7.68 mmol) of 5-(2,4-bis(methoxymethoxy)-5-isopropylphenyl)-4-(4-methoxyphenyl)-4H-[1,2,4]triazol-3-one (F60-03: anintermediate of Example 2-7) was added to a suspension of tribromoboranedimethylsulfide complex (11.65 g, 37.26 mol) and 1,2-dichloroethane (250mL) in 300 mL eggplant shaped flask and stirred at room temperature for2 hours and further at an external temperature of 80° C. for 24 hours.After completing the reaction, the reaction mixture was returned to roomtemperature and mixed with n-hexane (250 mL). The precipitates werecollected by filtration, washed with diethyl ether and then purified byDIAION HP-20 column chromatography (water-methanol gradient elution),followed by CHP-20 column chromatography (water-methanol gradientelution) to obtain the title compound (OH-all: 370 mg, 14.8%). Thefractions which were not sufficiently separated and purified werepurified by HPLC fractionation to obtain the title compound (OH-all: 300mg, total 670 mg, 26.8%).

LC/MS (Method 3): m/z (ESI, POS): 328 [M+H]⁺; retention time: 4.13minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 11.84 (1H, brs), 9.65 (1H, brs),9.59 (1H, brs), 9.44 (1H, brs), 6.98 (2H, d, J=8.8 Hz), 6.77 (1H, s),6.73 (2H, d, J=8.8 Hz), 6.26 (1H, s), 2.97 (1H, m), 0.96 (6H, d, J=7.0Hz).

Example 2-11 Preparation of4-[3-(2,4-dihydroxy-5-isopropylphenyl)-5-oxo-1,5-dihydro-[1,2,4]triazol-4-yl]-benzoicacid (OH-a12)

25% hydrogen bromide-acetic acid (3.5 mL) was added to an intermediateof Example 2-6,4-[3-(2,4-bis-allyloxy-5-isopropylphenyl)-5-oxo-1,5-dihydro-[1,2,4]triazol-4-yl]-benzoicacid (F61-03, 25 mg, 57 μmol) and stirred at room temperature for 1.5hours and then at 45° C. overnight. The solvent was distilled off tosome extent under reduced pressure, and azeotropic distillation wascarried out after adding toluene and acetonitrile. The crude productthus obtained was purified by silica gel column chromatography(chloroform, methanol) to obtain4-[3-(2,4-dihydroxy-5-isopropylphenyl)-5-oxo-1,5-dihydro-[1,2,4]triazol-4-yl]-benzoate(OH-a12: 11.8 mg, 58%).

LC/MS (Method 3): m/z (ESI, POS): 356 [M+H]⁺; retention time: 4.09minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=3:1) δ 0.86 (d, J=6.8 Hz, 6H), 3.02 (sept.,J=6.8 Hz, 1H), 6.33 (s, 1H), 6.61 (s, 1H), 7.39 (d, J=8.6 Hz, 2H), 8.14(d, J=8.6 Hz, 2H).

Example 2-12 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[2-(morpholin-4-yl)-pyrimidin-5-yl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a17)

The First Step: Preparation of G06-02

Under argon atmosphere, while stirring in an ice bath, phenylchlorothiono-formate (0.180 mL, 231 mg, 1.34 mmol) was added to adichloromethane (5 mL) solution of G06-01 [Heterocycles Vol. 6 (No. 12),1999-2004 (1977); 161 mg, 0.893 mmol] and4-(dimethyl-amino)-benzonitrile (261 mg, 1.79 mmol) and the mixture wasstirred for 1 hour under ice cold conditions. The reaction mixture wasmixed with saturated sodium chloride solution (5 mL) and 5% aqueoussodium hydrogencarbonate solution (5 mL) and extracted with ethylacetate. The ethyl acetate extracts were combined, washed with saturatedsodium chloride solution and dried over anhydrous sodium sulfate. Sodiumsulfate was removed by filtration, and the solvent was distilled offunder reduced pressure. Brown solids thus obtained were purified bycolumn chromatography (silica gel, dichloromethane-methanol) to obtainthe title compound (G06-02: yellow solids, 261 mg, 92%).

LC/MS (Method 3): m/z (ESI, POS): 317 [M+H]⁺; retention time: 5.82minutes.

The Second Step: Preparation of G06-03

Under argon atmosphere, while stirring at room temperature, IM4-a (271mg, 0.908 mmol) was added to a dimethylformamide (3 mL) solution ofG06-02 [261 mg, 0.825 mmol] and the mixture was stirred for 45 minuteswhile heating at a temperature of 90° C. The reaction mixture was mixedwith saturated sodium chloride solution and extracted with ethylacetate. The ethyl acetate extracts were combined, washed with saturatedsodium chloride solution and dried over anhydrous sodium sulfate. Sodiumsulfate was removed by filtration, and the solvent was distilled offunder reduced pressure. Yellow solids thus obtained were washed withether-hexane to obtain the title compound (G06-03: white solids, 388 mg,90%).

LC/MS (Method 3): m/z (ESI, POS): 521 [M+H]⁺, 543 [M+Na]⁺; retentiontime: 5.97 minutes.

NMR [400 MHz, CDCl₃, TMS] ppm: 1.000 (6H, d, J=7.0 Hz), 3.134 (1H,sept., J=7.0 Hz), 3.492 (3H, s), 3.580 (3H, s), 3.74-3.82 (8H, m), 5.252(2H, s), 5.468 (2H, s), 7.004 (1H, s), 7.743 (1H, s), 8.365 (2H, s),9.153 (1H, bs), 11.26 (1H, b), 12.06 (1H, b).

The Third Step: Preparation of G06-04

A mixture of G06-03 (380 mg, 0.73 mmol) and 1.25M aqueous sodiumhydroxide (10 mL, 12.5 mmol) was stirred while heating at 80° C. for 1.5hours. Under ice cold conditions, the reaction mixture was mixed with 1Maqueous potassium bisulfite and aqueous sodium hydrogencarbonate forneutralization, and extracted with ethyl acetate. The ethyl acetateextracts were combined, washed with saturated sodium chloride solutionand dried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration, and the solvent was distilled off under reduced pressure.Yellow solids thus obtained were purified by column chromatography(silica gel, hexane-ethyl acetate) to obtain the title compound (G06-04:colorless foamy compound, 220 mg, 60%).

LC/MS (Method 3): m/z (ESI, POS): 503 [M+H]⁺, 525 [M+Na]⁺; retentiontime: 6.15 minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.181 (6H, d, J=7.0 Hz), 3.232 (1H,sept., J=7.0 Hz), 3.271 (3H, s), 3.487 (3H, s), 3.73-3.83 (8H, m), 4.906(2H, s), 5.196 (2H, s), 6.861 (1H, s), 7.217 (1H, s), 8.227 (2H, s),11.745 (1H, s).

The Fourth Step: Preparation of G06-05

Under argon atmosphere, while stirring at room temperature, methyliodide (0.026 mL, 59 mg, 0.414 mmol) was added to a mixture of G06-04(208 mg, 0.414 mmol), solid potassium carbonate (57 mg, 0.414 mmol),ethanol (3 mL) and tetrahydrofuran (1.5 mL), and stirring was continuedat room temperature for 40 minutes. Under ice cold conditions, thereaction mixture was mixed with aqueous ammonium chloride and extractedwith ethyl acetate. The ethyl acetate extracts were combined, washedwith saturated sodium chloride solution and dried over anhydrous sodiumsulfate. Sodium sulfate was removed by filtration, and the solvent wasdistilled off under reduced pressure. The colorless syrup thus obtainedwas purified by column chromatography (silica gel, 1. hexane-ethylacetate; 2. ethyl acetate-methanol) to obtain the title compound(G06-05: colorless foam, 193 mg, 90%).

LC/MS (Method 3): m/z (ESI, POS): 517 [M+H]⁺, 539 [M+Na]⁺; retentiontime: 6.29 minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.185 (6H, d, J=7.0 Hz), 2.745 (3H,s), 3.228 (1H, sept., J=7.0 Hz), 3.247 (3H, s), 3.480 (3H, s), 3.73-3.83(8H, m), 4.896 (2H, s), 5.179 (2H, s), 6.823 (1H, s), 7.326 (1H, s),8.117 (2H, s).

The Fifth Step: Preparation of G06-06 (sulfone) and G-06-07 (sulfoxide)

Under argon atmosphere, while stirring under ice cold conditions,3-chloroperbenzoic acid (65 mg, 0.38 mmol) was added to adichloromethane (3 mL) solution of G06-05 (191 mg, 0.37 mmol) and themixture was stirred under ice cold conditions for 50 minutes. Next,while stirring under ice cold conditions, additional 3-chloroperbenzoicacid (27 mg, 0.15 mmol) was added and the mixture was stirred for 70minutes under ice cold conditions. Further, an addiitonal3-chloroperbenzoic acid (36 mg, 0.21 mmol) was added and the mixture wasstirred for 1 hour and 40 minutes under ice cold conditions, and 3 hoursat room temperature. Under ice cold conditions, an aqueous solution (5mL) of sodium sulfite (126 mg, 1 mmol) was added to the ice-coldreaction mixture and the mixture was stirred for 10 minutes. Finally,the reaction mixture was made alkaline by adding sodiumhydrogencarbonate and extracted with dichloromethane. Thedichloromethane extracts were combined, dried over anhydrous sodiumsulfate. Sodium sulfate was filtered off and the solvents of thefiltrate were distilled off under reduced pressure and a colorless syrupwas obtained, which was purified by column chromatography (silicagel, 1. hexane-ethyl acetate; 2. ethyl acetate-methanol) to obtain thetitle sulfone (G06-06: colorless foam, 119 mg, 59%) and the titlesulfoxide (G06-07: colorless foam, 39 mg, 20%). G06-06 (sulfone)

LC/MS (Method 3): m/z (ESI, POS): 549 [M+H]⁺, 571 [M+Na]⁺; retentiontime: 6.20 minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.189 (6H, d, J=7.0 Hz), 3.238 (1H,sept., J=7.0 Hz), 3.279 (3H, s), 3.488 (3H, s), 3.565 (3H, s), 3.742(4H, t, J=4.6 Hz), 3.814 (4H, t, J=4.6 Hz), 4.937 (2H, s), 5.199 (2H,s), 6.867 (1H, s), 7.296 (1H, s), 8.226 (2H, s). G06-07 (sulfoxide)

LC/MS (Method 3): m/z (ESI, POS): 533 [M+H]⁺; retention time: 5.62minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.191 (6H, d, J=7.0 Hz), 3.241 (1H,sept., J=7.0 Hz), 3.260 (3H, s), 3.320 (3H, s), 3.489 (3H, s), 3.73-3.77(4H, m), 3.80-3.85 (4H, m), 4.917 (2H, s), 5.198 (2H, s), 6.865 (1H, s),7.318 (1H, s), 8.237 (2H, s).

The Sixth Step: Preparation of G06-08

While stirring at room temperature, 1.25M aqueous sodium hydroxide (0.84mL, 1.05 mmol) was added to a dimethylsulfoxide (0,84 mL) solution ofG06-06 (sulfone, 115 mg, 0.21 mmol) and the mixture was heated at 70° C.for 40 minutes while stirring. After completing the reaction, an aqueoussolution of ammonium chloride was added to the reaction mixture underice cold conditions, and the reaction mixture was extracted with ethylacetate. The ethyl acetate extracts were combined, washed with sodiumchloride solution and then dried over anhydrous sodium sulfate. Sodiumsulfate was removed by filtration, and the solvent was distilled offunder reduced pressure, and a colorless syrup was obtained, which waspurified by column chromatography (silica gel, hexane-ethyl acetate) toobtain the title compound (G06-08, colorless foam, 89 mg, 87%).

LC/MS (Method 3): m/z (ESI, POS): 487 [M+H]⁺; retention time: 5.605minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.194 (6H, d, J=7.0 Hz), 3.240 (1H,sept., J=7.0 Hz), 3.245 (3H, s), 3.486 (3H, s), 3.710-3.755 (4H, m),3.755-3.800 (4H, m), 4.884 (2H, s), 5.190 (2H, s), 6.850 (1H, s), 7.254(1H, s), 8.163 (2H, s), 9.521 (1H, s).

The Seventh Step: Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[2-(morpholin4-yl)-pyrimidin-5-yl]-2,4-dihydro-[1,2,4]triazol-3-one (OH-a17)

Under argon atmosphere, while stirring in ice cold conditions, a1,4-dioxane solution of hydrochloric acid (4 N, 0.85 mL, 3.4 mmol) wasadded to a methanol (0.85 mL) solution of G06-08 (85 mg, 0.175 mmol) andstirred at room temperature for 3 hours. After completing the reaction,the reaction mixture was poured into a cold 5% aqueous sodiumhydrogencarbonate (10 mL)-saturated sodium chloride solution, and themixture was extracted with ethyl acetate. The ethyl acetate extractswere combined, washed with sodium chloride solution and dried overanhydrous sodium sulfate. Sodium sulfate was removed by filtration, thesolvent was distilled off under reduced pressure, and thus obtainedwhite solids were washed with ether-hexane to obtain the title compound(OH-a17: white solids, 53 mg, 76%).

LC/MS (Method 3): m/z (ESI, POS): 399 [M+H]⁺, 421 [M+Na]⁺; retentiontime: 4.400 minutes.

¹H-NMR [400 MHz, DMSO-d₆, TMS] ppm: 1.071 (6H, d, J=7.0 Hz), 3.022 (1H,sept., J=7.0 Hz), 3.60-3.68 (8H, m), 6.248 (1H, s), 6.984 (1H, s), 8.150(2H, s), 9.40-9.70 (2H, b), 11.94 (1H, b).

Example 2-13 Synthesis of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-isopropyl-2,4-dihydro-[1,2,4]triazol-3-one(OH-a21)

The First Step: Synthesis of F63-01

While stirring in ice cold conditions, isopropylisocyanate(2-isothiocyanato-propane) (3.0 mL, 28.2 mmol) was added to an ethanol(10 mL) solution of hydrazine monohydrate (2.88 g, 56.4 mmol). Afterstirring at room temperature for 30 minutes, the solvent was distilledoff under reduced pressure. The residue was extracted with chloroform,and the extract was washed twice with water and then with saturatedsodium chloride solution. After drying over anhydrous sodium, sodiumsulfate was removed by filtration and the solvent was distilled offunder reduced pressure. Solids thus obtained were dried under reducedpressure to obtain F-63-01 (white solids, 3.59 g, 95.8%).

LC/MS (Method 3): m/z (ESI, POS): 134 [M+H]⁺; retention time: 2.19minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.26 (d, J=6.6 Hz, 6H), 3.71 (s, 2H),4.49-4.58 (m, 1H), 7.14 (bs, 1H), 7.25 (br, 1H).

The Second Step: Synthesis of F63-02

At room temperature, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride n hydrate (DMT-MM, 2.34 g) was added to adimethylformamide (10 mL) solution of5-isopropyl-2,4-bis-methoxymethoxy-benzoic acid (2.00 g, 7.05 mmol) andF63-01 (984.0 mg, 7.39 mmol) obtained in the first step and stirred.After 3 hours, DMT-MM (250.4 mg) was further added and stirred for 1hour, and then the reaction was stopped by adding water. The reactionmixture was neutralized by adding an aqueous solution of saturatedsodium hydrogencarbonate and extracted with ethyl acetate. The extractwas washed with water and then saturated sodium chloride solution. Afterdrying the extract over anhydrous sodium, sodium sulfate was removed byfiltration, and the solvent was distilled off under reduced pressure.The solids thus obtained were dried under reduced pressure to obtainF63-02 (white solids, 2.82 g, 100%).

LC/MS (Method 3): m/z (ESI, POS): 400 [M+H]⁺; retention time: 6.32minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.23 (d, J=7.0 Hz, 6H), 3.26 (sept., J=7.0 Hz,1H), 3.51 (s, 3H), 3.58 (s, 3H), 4.40-4.49 (m, 1H), 5.27 (s, 2H), 5.43(s, 2H), 6.99 (s, 1H), 7.97 (s, 1H).

The Third Step:4-isopropyl-5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(F63-03)

At room temperature, 10% aqueous potassium hydroxide (20 mL) and 5%potassium hydroxide-ethanol solution (5 mL) were added to F63-02 (2.81g, 7.04 mmol) obtained in the second step, and the mixture was stirredat 90° C. for 13 hours. After returned to room temperature, chloroformand saturated sodium chloride solution were added and stirred for awhile. After this the reaction mixture was extracted with chloroform,and the extract was washed with saturated sodium chloride solution.After drying over anhydrous sodium, sodium sulfate was removed byfiltration, and the solvent was distilled off under reduced pressure toobtain an orange colored foam. After suspension purification inhexane-ethyl acetate (2:1), the solids were collected by filtration anddried under reduced pressure. After evaporating the solvent from thefiltrate under reduced pressure, the residue was purified by silica gelcolumn chromatography (hexane, ethyl acetate). Together,4-isopropyl-5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(F63-03, white solids, 1.96 g, 73%) was obtained.

LC/MS (Method 3): m/z (ESI, POS): 382 [M+H]⁺; retention time: 6.59minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.20 (d, J=7.0 Hz, 6H), 1.50 (d, J=7.0 Hz,6H), 3.29 (sept., J=7.0 Hz, 1H), 3.41 (s, 3H), 3.52 (s, 3H), 4.62(sept., J=7.0 Hz, 1H), 5.12 (s, 2H), 5.26 (s, 2H), 7.00 (s, 1H), 7.10(s, 1H), 10.53 (s, 1H).

The Fourth Step:4-isopropyl-3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(F63-04)

4-isopropyl-5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(F63-03, 1.96 g, 5.14 mmol) and potassium carbonate (710.4 mg, 5.14mmol) were weighed and placed in a reaction vessel, and ethanol (30 mL)was added and then methyl iodide (0.32 mL, 5.14 mmol) was added. Afterheating at 80° C. for 1 hour while stirring, the mixture was returned toroom temperature, and the solvent was distilled off under reducedpressure. After adding water, the reaction system was extracted withethyl acetate, and the extract was washed with saturated sodium chloridesolution. After drying the extract over anhydrous sodium, sodium sulfatewas removed by filtration and the solvent was distilled off underreduced pressure to obtain yellow syrup. After purification by silicagel column chromatography (chloroform, methanol),4-isopropyl-3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-5-methylsulfanyl-4H-[1,2,4]triazole (F63-04, pale yellowsyrup, 1.93 g, 95%) was obtained.

LC/MS (Method 3): m/z (ESI, POS): 396 [M+H]⁺; retention time: 6.49minutes.

The Fifth Step:4-isopropyl-3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-5-methylsulfonyl-4H-[1,2,4]triazole(F63-05)

While stirring in ice cold conditions, 3-chloroperbenzoic acid (4.20 g,24.4 mmol) was added to a methylene chloride (10 mL) solution of4-isopropyl-3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(F63-04, 1.93 g, 4.87 mmol). After stirring at room temperature for 2hours, the mixture was bring back to ice cold conditions again, andsaturated aqueous sodium thiosulfate and then saturated aqueous sodiumhydrogencarbonate were added. After stirring for a while, the reactionmixture was extracted with ethyl acetate, and the extract was washedwith saturated sodium chloride solution. After drying the extract overanhydrous sodium, sodium sulfate was removed by filtration and thesolvent was distilled off under reduced pressure to obtain a pale yellowfoam. After purification by silica gel column chromatography (hexane,ethyl acetate),4-isopropyl-3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-5-methylsulfonyl-4H-[1,2,4]triazole(F63-05, colorless foam, 1.65 g, 79%) was obtained.

LC/MS (Method 3): m/z (ESI, POS): 428 [M+H]⁺; retention time: 6.56minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.20 (d, J=7.0 Hz, 6H), 1.51 (d, J=7.0 Hz,6H), 3.30 (sept., J=7.0 Hz, 1H), 3.39 (s, 3H), 3.53 (s, 3H), 3.63 (s,3H), 4.69 (sept., J=7.0 Hz, 1H), 5.11 (s, 2H), 5.27 (s, 2H), 7.02 (s,1H), 7.16 (s, 1H).

The Sixth Step:4-isopropyl-5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-2,4-dihydro-[1,2,4]triazol-3-one(F63-06)

At room temperature, aqueous sodium hydroxide (1.0 M aqueous solution, 9mL) was added to a dimethylsulfoxide (11 mL) solution of4-isopropyl-3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-5-methylsulfonyl-4H-[1,2,4]triazole (F63-05, 1.63 g, 3.82 mmol). The mixture washeated at 90-100° C. for 12 hours while stirring. After returning toroom temperature, water was added. The reaction mixture was extractedwith ethyl acetate, and the extract was washed twice with water and thenwith saturated sodium chloride solution. After drying the extract overanhydrous sodium, sodium sulfate was removed by filtration and thesolvent was distilled off under reduced pressure to obtain white solids.After suspension purification by diethyl ether, the solids obtained byfiltration were dried under reduced pressure to obtain4-isopropyl-5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-2,4-dihydro-[1,2,4]triazol-3-one(F63-06, white solids, 1.09 g, 78%).

LC/MS (Method 3): m/z (ESI, POS): 366 [M+H]⁺; retention time: 5.88minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.21 (d, J=7.0 Hz, 6H), 1.46 (d, J=7.0 Hz,6H), 3.28 (sept., J=7.0 Hz, 1H), 3.43 (s, 3H), 3.52 (s, 3H), 3.93(sept., J=7.0 Hz, 1H), 5.14 (s, 2H), 5.25 (s, 2H), 6.99 (s, 1H), 7.16(s, 1H), 8.93 (s, 1H).

The Seventh Step:5-(2,4-dihydroxy-5-isopropylphenyl)-4-isopropyl-2,4-dihydro-[1,2,4]triazol-3-one(OH-a21)

5 N hydrochloric acid (7 mL) was added to a methanol (13 mL) solution of4-isopropyl-5-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-2,4-dihydro-[1,2,4]triazol-3-one(F63-06, 1.03 g, 2.82 mmol) and the mixture was stirred overnight. Aftercooling to ice cold conditions, the reaction mixture was neutralized byadding saturated aqueous sodium hydrogencarbonate. The solution wasextracted with ethyl acetate, and the extract was washed with water andthen with saturated sodium chloride solution. After drying the extractover anhydrous sodium, sodium sulfate was removed by filtration and thesolvent was distilled off under reduced pressure to obtain white solids.After crude purification by silica gel column chromatography(chloroform, methanol), the solids were further purified by silica gelcolumn chromatography (NHsilica, diethyl ether, methanol) to obtain5-(2,4-dihydroxy-5-isopropylphenyl)-4-isopropyl-2,4-dihydro-[1,2,4]triazol-3-one(OH-a21, white solids, 581 mg, 74%).

LC/MS (Method 3): m/z (ESI, POS): 278 [M+H]⁺; retention time: 4.38minutes.

FAB-MS: m/z (POS): 278 [M+H]⁺; melting point: 261-262° C. (dec.).

¹H-NMR (400 MHz, DMSO-d₆) δ 1.11 (d, J=7.0 Hz, 6H), 1.31 (d, J=6.8 Hz,6H), 3.08 (sept., J=7.0 Hz, 1H), 3.80 (sept., J=6.8 Hz, 1H), 6.45 (s,1H), 6.88 (s, 1H), 9.69 (bs, 2H), 11.47 (bs, 1H).

Example 2-14 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-piperidin-1-yl-2,4-dihydro-[1,2,4]triazol-3-one(OH-a24) trifluoroacetate

The First Step: Preparation of2,4-bis-benzyloxy-5-isopropyl-N-piperidin-1-yl-benzamide (F67-02)

2,4-bis-benzyloxy-5-isopropylbenzoic acid (F67-01: 188 mg, 0.5 mmol),dimethylformamide (2 mL) and 1-hydroxy-1,2,3-benzotriazole (72 mg, 0.55mmol) were placed in a test tube and then 1-amino-piperidine (0.059 mL,0.55 mmol) was added. A mixed solution of dimethylformamide (1 mL),1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide hydrochloride (105 mg,0.55 mmol) and triethylamine (0.15 mL, 1.1 mmol) was slowly added to thereaction mixture at 0° C. while stirring. Further, the reaction mixturewas allowed to come to room temperature and stirred for 24 hours. Aftercompleting the reaction, the reaction mixture was extracted with ethylacetate, and the extract was washed 4 times with saturated sodiumchloride, dried with sodium sulfate and then concentrated under reducedpressure. The residue was purified by silica gel column chromatography(methylene chloride/methanol) followed by silica gel columnchromatography (hexane/ethyl acetate) to obtain the title compound(F67-02: 198 mg, 86.4%).

LC/MS (Method 5): m/z (ESI, POS): 459 [M+H]⁺; retention time: 7.44minutes.

The Second Step: Preparation of2,4-bis-benzyloxy-5-isopropyl-N-piperidin-1-yl-thiobenzamide (F67-03)

2,4-bis-benzyloxy-5-isopropyl-N-piperidin-1-yl-benzamide (F67-02: 198mg, 0.43 mmol), toluene (5 mL) and Lawesson's reagent (157 mg, added intwo portions) were placed in a test tube and heated for 2.5 hours underreflux. After the reaction was completed, the reaction mixture wasconcentrated under reduced pressure. The foam thus obtained wassubjected to the next reaction without further purification.

LC/MS (Method 5): m/z (ESI, POS): 475 [M+H]⁺; retention time: 7.86minutes.

The Third Step: Preparation of2,4-bis-benzyloxy-5-isopropyl-N-piperidin-1-yl-benzene-carbohydrazonamide(F67-04)

2,4-bis-benzyloxy-5-isopropyl-N-piperidin-1-yl-thiobenzamide (unpurifiedproduct of the previous step: F67-03), ethanol (5 mL), hydrazinemonohydrate (0.5 mL) was placed in a test tube and heated for 1.5 hoursunder reflux. After completing the reaction, the reaction mixture wasconcentrated under reduced pressure while adding toluene several times.The residue thus obtained was subjected to the next step withoutpurification.

LC/MS (Method 5): m/z (ESI, POS): 473 [M+H]⁺; retention time: 3.51minutes.

The Fourth Step: Preparation of5-(2,4-bis-benzyloxy-5-isopropyl-phenyl)-4-piperidin-1-yl-2,4-dihydro-[1,2,4]triazol-3-thione(F67-05)

2,4-bis-benzyloxy-5-isopropyl-N-piperidin-1-yl-benzene-carbohydrazonamide(unpurified product of the previous step: F67-04), tetrahydrofuran (3mL) and triphosgene (42 mg) were placed in a test tube and stirred atroom temperature. After completing the reaction, methanol and sodiumhydrogencarbonate were added and the mixture was stirred for a while,and then solids were separated by filtration. The mother liquor thusobtained was concentrated under reduced pressure, and the residue waspurified by silica gel column chromatography (hexane/ethyl acetate) toobtain the title compound (F67-05: 92 mg, 36.7%).

LC/MS (Method 5): m/z (ESI, POS): 499 [M+H]⁺; retention time: 7.44minutes.

The Fifth Step: Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-piperidin-1-yl-2,4-dihydro-[1,2,4]triazol-3-one(OH-a24) trifluoroacetate

5-(2,4-bis-benzyloxy-5-isopropyl-phenyl)-4-piperidin-1-yl-2,4-dihydro-[1,2,4]triazol-3-thione(F67-05: 92 mg, 0.18 mmol), methylene chloride (2 mL) and a methylenechloride solution of boron trichloride (1 mol/L, 1 mL) were placed in atest tube and stirred at room temperature for 1 hour. After completingthe reaction, methanol and sodium hydrogencarbonate were added to thereaction mixture and solids were separated by filtration. The motherliquor thus obtained was concentrated under reduced pressure and theresidue was purified by HPLC fractionation to obtain the title compound(OH-a24 trifluoroacetate, 9.5 mg, 16.6%).

LC/MS (Method 3): m/z (ESI, POS): 319 [M+H]⁺; retention time: 5.74minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 11.8 (1H, s), 9.94 (1H, s), 9.75(1H, s), 7.74 (1H, s), 6.38 (1H, s), 3.70-3.45 (2H, brs), 3.12 (1H,sept, J=6.6 Hz), 3.20-2.90 (2H, brs), 1.80-1.40 (6H, brs), 1.15 (6H, d,J=6.6 Hz)

Example 2-15 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(2-morpholin-4-yl-ethyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a26) monohydrochloride

The title compound (OH-a26monohydrochloride) was synthesized in thesimilar manner to that of Example 2-16 using(2-morpholin-4-yl-ethyl)-thiocarbamic acid O-phenyl ester in place ofF93-06 of Example 2-16.

LC/MS (Method 6): m/z (ESI, POS): 349 [M−HCl+H]⁺; retention time: 3.86minutes.

¹H-NMR (400 MHz, DMSO-d₆) δ 1.13 (d, J=7.0 Hz, 6H), 2.96-3.14 (m, 3H),3.59-3.74 (m, 2H), 4.10 (t, J=6.1 Hz, 2H), 3.94 (bd, J=12.3 Hz, 2H),6.56 (s, 1H), 7.00 (s, 1H), 9.86 (s, 1H), 10.03 (s, 1H), 10.32 (br, 1H),11.92 (s, 1H).

Example 2-16 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[3-(morpholin-4-yl)-propyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a27)

The First Step: Preparation of F93-01 (IM4-a)

IM3-a (1137 mg, 4 mmol), hydrazine•monohydrate (240 mg, 4.8 mmol) anddimethylformamide (15 mL) were placed in a 100 mL flask, and to this1-hydroxy benztriazolemonohydrate (656 mg, 4.8 mmol) and1-ethyl-3-(3′-dimethylaminopropyl)carbodiimidehydrochloride (932 mg,4.86 mmol) were added under ice cold conditions, and the mixture wasstirred for 20 hours. After adding water (100 mL) and saturated aqueoussodium hydrogencarbonate (15 mL), the reaction mixture was extractedtwice with ethyl acetate (100 mL). The organic layer was washed withsaturated sodium chloride solution, dried with sodium sulfate and thenconcentrated under reduced pressure to obtain the title crude compound(F93-01: 1302 mg).

LC/MS (Method 3): m/z (ESI, POS): 299 [M+H]⁺; retention time: 4.69minutes.

The Second Step: Preparation of F93-02

Crude F93-01 (298 mg, 1.0 mmol), F93-06 (280 mg, 1.5 mmol) and ethanol(15 mL) were placed in a 50 mL flask and heated for 2 hours underreflux, and then the reaction mixture was concentrated under reducedpressure to obtain the crude title compound (F93-02).

LC/MS (Method 3): m/z (ESI, POS): 485 [M+H]⁺; retention time: 3.36minutes.

Preparation of F93-06 3-morpholinopropylamine (288.4 mg, 2 mmol) andanhydrous dichloromethane (10 mL) were placed in a 100 mL flask, and tothis a mixture of O-phenyl chlorothionoformate (332 μL, 2.4 mmol) andpyridine (232 μL, 2.88 mmol) under ice cold conditions and stirred for 2hours. The reaction mixture was mixed with water (10 mL) and saturatedaqueous sodium hydrogencarbonate (5 mL), and extracted twice withchloroform (15 mL). After washing with saturated sodium chloridesolution, the organic layer was dried with sodium sulfate andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (chloroform:methanol=10:1) to obtain the titlecompound (F93-06: 425 mg, yield 75.7%).

LC/MS (Method 3): m/z (ESI, POS): 281 [M+H]⁺; retention time: 1.08minutes.

The Third Step: Preparation of F93-03

The crude product produced in the second step (F93-02, 465 mg) wasdissolved in 10% aqueous potassium hydroxide (12 mL) and 5% ethanolsolution of potassium hydroxide (6 mL) and heated for 1.5 hours underreflux. After concentrating the reaction mixture, the residue was mixedwith water (30 mL) and extracted twice with ethyl acetate (30 mL). Afterconcentrating the extract under reduced pressure, the residue waspurified by silica gel column chromatography (chloroform:methanol=20:1)to obtain the title compound (F93-03: 247 mg, yield 52.0%, in total 3steps)

LC/MS (Method 3): m/z (ESI, POS): 467 [M+H]⁺; retention time: 3.30minutes.

The Fourth Step: Preparation of F93-04

F93-03 (247 mg, 0.52 mmol) dissolved in ethanol (8 mL) was mixed withpotassium carbonate (72 mg, 0.52 mmol) and methyl iodide (33 μL, 0.52mmol) and heated for 1 hour under reflux. After concentrating, thereaction mixture was mixed with water (10 mL) and extracted twice withethyl acetate (15 mL). The organic layer was concentrated under reducedpressure to obtain the title crude compound (F93-04, 234 mg).

LC/MS (Method 3): m/z (ESI, POS): 481 [M+H]⁺; retention time: 3.11minutes.

The Fifth Step: Preparation of F93-05

After dissolving the crude F93-04 (234 mg) in dichloromethane (10 mL),m-chloroperbenzoate (345 mg, 2 mmol) was added to it and the mixture wasstirred for 20 hours. The reaction mixture was mixed with chloroform (20mL) and 10% potassium bisulfite solution (30 mL), stirred for 10minutes, and then the organic layer was separated. The organic layer waswashed with saturated sodium chloride solution, dried with sodiumsulfate and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (chloroform:methanol=20:1)to obtain the title compound (F93-05: 147 mg, yield 55.1%, in total 2steps).

LC/MS (Method 3): m/z (ESI, POS): 513 [M+H]⁺; retention time: 3.40minutes.

The Sixth Step: Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[3-(morpholin-4-yl)-propyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a27)

After dissolving F93-05 (91 mg, 0.177 mmol) in dimethylsulfoxide (0.5mL), 3 N aqueous sodium hydroxide (0.5 mL) was added to it and stirredat 90° C. for 1.5 hours. After adding water (15 mL), the reactionmixture was extracted twice with ethyl acetate (15 mL). The organiclayer was washed with saturated sodium chloride solution, dried withsodium sulfate and then concentrated under reduced pressure. The residuewas dissolved in methanol (2 mL) and mixed with 5 N hydrochloric acid(1.5 mL), and the mixture was stirred at 55° C. for 1 hour. Afterconcentrating, the residue was dissolved in methanol (6 mL) and mixedwith silica gel (350 mg) and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography(chloroform:methanol=10:1-3:1) to obtain the title compound (OH-a27: 53mg, yield 82.6%).

LC/MS (Method 4): m/z (ESI, POS): 363 [M+H]⁺; retention time: 6.42minutes.

¹H-NMR (400 MHz, CD₃OD, ppm): 7.06 (1H, s), 6.43 (1H, s), 4.61 (2H,brs), 3.78 (2H, brs), 3.75 (4H, brs), 3.18 (1H, m), 2.86 (4H, brs), 1.98(2H, m), 1.19 (6H, d, J=6.95)

Example 2-17 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-[3-(2-oxo-pyrrolidin-1-yl)propyl]-2,4-dihydro-[1,2,4]triazol-3-one(OH-a30)

N-(3-isothiocyanatopropyl)-2-pyrrolidinone was obtained by the similarprocess to that of Example 2-2(B) in 1 step usingN-(3-aminopropyl)-2-pyrrolidinone in place of4-morpholin-4-ylmethyl-phenylamine (F45-000) in Example 2-2 (B). Thetitle compound (OH-a30) was obtained using this compound in the similarprocess to that of Example 2-13 in 7 steps.

LC/MS (Method 1): m/z (ESI, POS): 361 [M+H]⁺; retention time: 4.10minutes.

¹H-NMR [400 MHz, CDCl₃+CD₃OD (three drops)] δ 1.20 (d, J=7.0 Hz, 6H),1.82-2.00 (m, 4H), 2.34 (t, J=7.9 Hz, 2H), 3.14-3.28 (m, 5H), 3.62-3.70(m, 2H), 6.40 (s, 1H), 7.30 (s, 1H).

Example 2-18 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(2-methoxy-ethyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a32)

The bis(methoxymethyl) protected compound of the title compound (OH-a32)was obtained from the compound of Example 3-14 (SFN-a32) protected by abis(methoxymethyl) group by the similar manner to that of the sixth stepof Example 2-12. This compound was deprotected in the similar manner tothat in the seventh step of Example 1-1 to obtain the title compound(OH-a32).

LC/MS (Method 3): m/z (ESI, POS): 294 [M+H]⁺; retention time: 3.99minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=3:1, ppm): 7.25 (1H, s), 6.39 (1H, s), 3.91(2H, t, J=5.86 Hz), 3.57 (2H, t, J=5.86 Hz), 3.24 (3H, s), 3.23 (1H, m),1.20 (6H, d, J=6.96 Hz)

Example 2-19 Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(2-hydroxy-1-hydroxymethyl-ethyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a33)

The First Step: Preparation of F77-02

The title compound was synthesized from 5-amino-2,2-dimethyl-1,3-dioxane(F77-01) that is known in the literature (Chem. Pharm. Bull. Vol. 44,No. 12, 2205-2212, 1996) by the similar manner to that of the first stepof Example 2-12.

LC/MS (Method 3): m/z (ESI, POS): 268 [M+H]⁺, 210; retention time: 6.007minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.464 (3H, s), 1.532 (3H, s),3.875-4.050 (2H, m), 4.150-4.270 (3H, m), 7.07-7.13 (2H, m), 7.21-7.36(1H, m), 7.38-7.47 (2H, m), 7.595 (1H, d, J=7.7 Hz).

The Second Step; Preparation of F77-03

Under argon atmosphere, a dimethylformamide (2 mL) solution of IM4-a(180 mg, 0.603 mmol) and F77-02 [161 mg, 0.603 mmol] was heated at 100°C. for 1.5 hours on an oil bath while stirring. The reaction mixture wasmixed with sodium chloride solution and extracted with ethyl acetate.The ethyl acetate extracts were combined, washed with sodium chloridesolution and dried over anhydrous sodium sulfate. Sodium sulfate wasremoved by filtration, the solvent was distilled off under reducedpressure and the yellow syrup thus obtained was purified by columnchromatography (silica gel, hexane-ethyl acetate) to obtain the titlecompound (F77-03: colorless foam, 194 mg, 68%).

LC/MS (Method 3): m/z (ESI, POS): 472 [M+H]⁺, 494 [M+Na]⁺,414; retentiontime: 6.31 minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.220 (6H, d, J=6.9 Hz), 1.446 (3H,s), 1.641 (3H, s), 3.243 (1H, sept., J=6.9 Hz), 3.499 (3H, s), 3.564(3H, s), 3.856 (2H, m), 4.143 (2H, m), 4.399 (1H, dt, Jd=8.3 Hz, Jt=2.5Hz), 5.256 (2H, s), 5.408 (2H, s), 6.967 (1H, s), 7.496 (1H, d, J=8.3Hz), 8.011 (1H, s), 8.00-11.5 (2H, b).

The Third Step: Preparation of F77-04

A mixture of F77-03 (165 mg, 0.35 mmol) and 1.25M aqueous sodiumhydroxide (5 mL, 6.25 mmol) was heated for 3 hours under reflux. Underice cold conditions, the reaction mixture was mixed with 1M aqueouspotassium bisulfate (6.25 mL) and extracted with ethyl acetate. Theethyl acetate extracts were combined, washed with saturated sodiumchloride solution, dried over anhydrous sodium sulfate. Sodium sulfatewas removed by filtration, the solvent was distilled off under reducedpressure, and the colorless syrup thus obtained was purified by columnchromatography (silica gel, hexane-ethyl acetate) to obtain the titlecompound (F77-04: colorless foam, 113 mg, 71%).

LC/MS (Method 3): m/z (ESI, POS): 454 [M+H]⁺, 396; retention time: 6.75minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.206 (6H, d, J=7.0 Hz), 1.370 (3H,s), 1.684 (3H, s), 3.276 (1H, sept., J=7.0 Hz), 3.435 (3H, s), 3.533(3H, s), 3.75 (2H, m), 4.311 (1H, m), 5.159 (2H, s), 5.281 (2H, s), 5.40(2H, m), 7.051 (1H, s), 7.133 (1H, s).

The Fourth Step: Preparation of F77-05

Under argon atmosphere, while stirring at room temperature, methyliodide (0.0182 mL, 42 mg, 0.293 mmol) was added to an ethanol (2 mL)suspension of F-77-04 (133 mg, 0.293 mmol) and solid potassiumcarbonate, and the mixture was stirred at room temperature for 2 hours.Under ice cold conditions, the reaction mixture was mixed with aqueousammonium chloride and extracted with ethyl acetate. The ethyl acetateextracts were combined, washed with saturated sodium chloride solution,dried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration and the solvent was distilled off under reduced pressure toobtain the title compound (F77-05: yellow foam, 125 mg, 91%).

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.206 (6H, d, J=7.0 Hz), 1.369 (3H,s), 1.573 (3H, s), 2.837 (3H, s), 3.275 (1H, sept., J=7.0 Hz), 3.394(3H, s), 3.526 (3H, s), 3.80 (2H, m), 4.340 (1H, tt, J=11.0, 5.5 Hz),4.510 (2H, t, J=11.0 Hz), 5.104 (2H, s), 5.267 (2H, s), 7.017 (1H, s),7.232 (1H, s).

The Fifth Step: Preparation of F77-06

Under argon atmosphere, while stirring under ice cold conditions,3-chloroperbenzoic acid (95 mg, 0.55 mmol) was added to adichloromethane (2 mL) solution of F77-05 (123 mg, 0.263 mmol) and themixture was stirred at room temperature for 3.5 hours. The reactionmixture was diluted by adding dichloromethane, and washed with aqueoussodium sulfite followed by sodium chloride solution, and then dried overanhydrous sodium sulfate. Sodium sulfate was removed by filtration andthe solvent was distilled off under reduced pressure, and the colorlessfoam thus obtained was purified by column chromatography (silica gel,hexane-ethyl acetate) to obtain the title compound (F77-06: colorlessfoam, 98 mg, 75%) G06-06 (sulfone)

LC/MS (Method 3): m/z (ESI, POS): 500 [M+H]⁺, 522 [M+Na]⁺; retentiontime: 6.74 minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.217 (6H, d, J=7.0 Hz), 1.353 (3H,s), 1.501 (3H, s), 3.296 (1H, sept., J=7.0 Hz), 3.412 (3H, s), 3.533(3H, s), 3.660 (3H, s), 3.84 (2H, m), 4.57-4.70 (3H, m), 5.138 (2H, s),5.289 (2H, s), 7.071 (1H, s), 7.208 (1H, s).

The Sixth Step: Preparation of F77-07

While stirring at room temperature, 1.25M aqueous sodium hydroxide (0.6mL, 0.75 mmol) was added to a dimethylsulfoxide (0.6 mL) solution ofF77-06 (75 mg, 0.15 mmol) and the mixture was heated at 90° C. for 1hour and 15 minutes while stirring. After completing the reaction, thereaction mixture was mixed with aqueous ammonium chloride under ice coldconditions and extracted with ethyl acetate. The ethyl acetate extractswere combined, washed with sodium chloride solution and then dried overanhydrous sodium sulfate. Sodium sulfate was removed by filtration, andthe solvent was distilled off under reduced pressure, and the colorlesssyrup was purified by column chromatography (silica gel, hexane-ethylacetate) to obtain the title compound (F77-07: colorless foam, 60 mg,92%).

LC/MS (Method 3): m/z (ESI, POS): 438 [M+H]⁺, 460 [M+Na]⁺, 380;retention time: 6.054 minutes.

¹H-NMR [400 MHz, CDCl₃, TMS] ppm: 1.210 (6H, d, J=6.9 Hz), 1.360 (3H,s), 1.635 (3H, s), 3.271 (1H, sept., J=6.9 Hz), 3.453 (3H, s), 3.531(3H, s), 3.69-3.80 (2H, m), 3.959 (1H, tt, J=11.2, 5.5 Hz), 4.816 (2H,t, J=11.2 Hz), 5.169 (2H, s), 5.269 (2H, s), 7.022 (1H, s), 7.161 (1H,s), 9.442 (1H, s).

The Seventh Step: Preparation of5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(2-hydroxy-1-hydroxymethyl-ethyl)-2,4-dihydro-[1,2,4]triazol-3-one(OH-a33)

While stirring under ice cold conditions, 6 N aqueous hydrochloric acid(0.5 mL, 3.0 mmol) was added to a methanol (1.0 mL) solution of F77-07(58 mg, 0.133 mmol), and the mixture was stirred at room temperature for17.5 hours. While stirring under ice cold conditions, 5% aqueous sodiumhydrogencarbonate (5 mL) and saturated sodium chloride were added to thereaction mixture, and the mixture was extracted with ethyl acetate. Theethyl acetate extracts were combined, washed with sodium chloridesolution and then dried over anhydrous sodium sulfate. Sodium sulfatewas removed by filtration, and the solvent was distilled off underreduced pressure, and the white solids thus obtained were washed withether to obtain the title compound (OH-a33: white solids, 33 mg, 80%).

LC/MS (Method 1): m/z (ESI, POS): 310 [M+H]⁺, 332 [M+Na]⁺; retentiontime: 2.54 minutes.

¹H-NMR [400 MHz, DMSO-d₆, TMS] ppm: 1.108 (6H, d, J=6.8 Hz), 3.067 (1H,sept., J=6.8 Hz), 3.60-3.80 (5H, m), 4.81 (2H, bs), 6.424 (1H, s), 6.963(1H, s), 9.50-9.70 (1H, b), 9.612 (1H, s), 11.624 (1H, s).

Example 3-1 Preparation of4-isopropyl-6-{5-methylsulfanyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-dioltrifluoroacetate (SMe-a02-TF)

The First Step: Preparation of4-{4-[3-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-5-methylsulfanyl-[1,2,4]triazol-4-yl]-benzyl}-morpholine(IM6-SMe-a02)

5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(IM6-S-a02: 292 mg, 0.57 mmol), potassium carbonate (78.6 mg, 0.57 mmol)and ethanol (5 mL) were placed in a 50 mL eggplant shaped flask, andthen methyl iodide (0.035 mL, 0.57 mL) was added, and the mixture washeated for 2 hours under reflux. The reaction mixture was filtered, andthe mother liquor was concentrated under reduced pressure, and theresidue thus obtained was purified by silica gel column chromatographyto obtain the title compound (IM6-SMe-a02: 206 mg, 39.1%).

LC/MS (Method 3): m/z (ESI, POS): 529 [M+H]⁺; retention time: 3.34minutes.

The Second Step: Preparation of4-isopropyl-6-{5-methylsulfanyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SMe-a02-TF)

4-{4-[3-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-5-methylsulfanyl-[1,2,4]traizol-4-yl]-benzyl}-morpholine(IM6-SMe-a02: 150 mg, 0.284 mmol), ethanol (1.5 mL) and then 5 Nhydrochloric acid (1.5 mL) were placed in a 50 mL eggplant shaped flask,and the mixture was stirred at room temperature for 24 hours. Aftercompleting the reaction, the reaction mixture was neutralized with 10 Naqueous sodium hydroxide and then extracted with ethyl acetate. Thecollected organic layer was dried with sodium sulfate and thenconcentrated under reduced pressure. The residue thus obtained waspurified by HPLC fractionation to obtain the title compound (SMe-a02-TF:32.6 mg, 20%).

LC/MS (Method 6): m/z (ESI, POS): 441 [M+H]⁺; retention time: 4.25minutes.

¹H-NMR (400 MHz, CD₃OD, TMS) ppm: 7.77 (2H, d, J=8.4 Hz), 7.61 (2H, d,J=8.4 Hz), 6.80 (1H, s), 6.36 (1H, s), 4.48 (2H, s), 4.10-3.90 (2H, br),3.90-3.60 (2H, br), 2.72 (1H, sept, J=6.4 Hz).

Example 3-2 Preparation of4-isopropyl-6-{5-methylsulfinyl-4-[4-(morpholin-4-oxid-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-dioltrifluoroacetate (SFX-a07-TF) and4-isopropyl-6-{5-methanesulfonyl-4-[4-(morpholin-4-oxid-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}benzene-1,3-diol(SFN-a07)

4-isopropyl-6-{5-methylsulfanyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-dioltrifluoroacetate (SMe-a02-TF: 41.4 mg, 0.093 mmol) and methylenechloride (2 mL) were placed in a test tube, and thenmetachloroperbenzoic acid (38 mg, 0.372 mmol) was added and the mixturewas stirred at room temperature for 24 hours. After competing thereaction, the reaction mixture was concentrated under reduced pressure,and the residue thus obtained was purified by HPLC fractionation toobtain the title compound (SFX-a07-TF: 20.6 mg, 37.8%) and the titlecompound (SFN-a07: 15.9 mg, 35%)

SFX-a07-TF

LC/MS (Method 6): m/z (ESI, POS): 473 [M+H]⁺; retention time: 3.99minutes.

MS (FAB, POS) m/z: 473 [M+H]⁺, 371 [M-morpholine N-oxide+H]⁺.

SFN-a07

LC/MS (Method 6): m/z (ESI, POS): 489 [M+H]⁺; retention time: 4.16minutes.

Example 3-3 Preparation of4-bromo-6-{5-methylsulfanyl-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-dioltrifluoroacetate (SMe-d01-TF)

The First Step: Preparation of4-{4-[3-(5-bromo-2,4-bis-methoxymethoxy-phenyl)-5-methylsulfanyl-[1,2,4]triazol-4-yl]-phenyl}-morpholine(IM6-SMe-d01)

5-(5-bromo-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-yl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-thione(Example 1-5, IM6-S-d01: 54 mg, 0.1 mmol), potassium carbonate (13.8 mg,0.17 mmol) and ethanol (5 mL) were placed in a test tube, and thenmethyl iodide (14.2 mg, 0.1 mmol) was added, and the mixture was heatedfor 2 hours under reflux. After completing the reaction, potassiumcarbonate was removed by filtration, and the mother liquor wasconcentrated. The crude product (IM6-SMe-d01) thus obtained wassubjected to the next reaction without further purification inparticular.

LC/MS (Method 3): m/z (ESI, POS): 551 [M+H]⁺; retention time: 5.86minutes.

The Second Step: Preparation of4-bromo-6-{5-methylsulfanyl-4-[4-(morpholin-4-yl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-dioltrifluoroacetate (SMe-d01-TF)

The crude product of4-{4-[3-(5-bromo-2,4-bis-methoxymethoxy-phenyl)-5-methylsulfanyl-[1,2,4]triazol-4-yl]-phenyl}-morpholine(IM6-SMe-d01), ethanol (1 mL) and then 5 N hydrochloric acid (1 mL) wereplaced in a test tube, and the mixture was stirred for 24 hours. Aftercompleting the reaction, the reaction mixture was neutralized with 10 Naqueous sodium hydroxide and then extracted with ethyl acetate. Thecollected organic layer was dried with sodium sulfate and thenconcentrated under reduced pressure. The residue thus obtained waspurified by HPLC fractionation to obtain the title compound (SMe-d01-TF:18.5 mg, 32%).

LC/MS (Method 3): m/z (ESI, POS): 461 [M+H]⁺; retention time: 5.48minutes.

¹H-NMR (400 MHz, CD₃OD, TMS) ppm: 7.23 (2H, d, J=9.2), 7.09 (2H, d,J=9.2), 6.94 (1H, s), 6.81 (1H, s), 4.00-3.95 (4H, br), 3.40-3.30 (4H,br), 2.70 (3H, s).

Example 3-4 (A) Preparation of4-isopropyl-6-{5-methanesulfonyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SFN-a02)

4-isopropyl-6-{5-methylsulfanyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-dioltrifluoroacetate (SMe-a02-TF: 56.8 mg, 0.129 mmol), methylene chloride(3 mL) and then metachloroperbenzoic acid (112 mg, 0.645 mmol) wereplaced in a test tube, and the mixture was stirred at room temperaturefor 24 hours. After completing the reaction, the reaction mixture wasconcentrated under reduced pressure. To the residue were added ethanol(3 ml) and a suspension of Raney nickel in ethanol (0.3 ml)successively, and the mixture was stirred at room temperature for 5.5hours in a hydrogen atmosphere.

After completing the reaction, the reaction mixture was filtered throughCelite, and the filtrate was concentrated under reduced pressure. Theresidue thus obtained was purified by preparative HPLC to obtain thetitle compound (SFN-a02: 3.7 mg, 6.1%).

LC/MS (Method 1): m/z (ESI, POS): 473 [M+H]⁺; retention time: 1.07minutes.

¹H-NMR (400 MHz, CDCL₃, TMS) ppm: 7.61 (2H, d, J=8.0), 7.46 (2H, d,J=8.0), 7.26 (1H, s), 6.53 (1H, s), 3.80-3.72 (4H, m), 3.64 (2H, s),3.50 (3H, s), 2.90 (1H, sept, J=6.8), 2.60-2.50 (4H, br), 0.75 (3H, s),0.73 (3H, s).

MS (FAB,POS) m/z: 473 [M+H]⁺, 387 [M-morpholine+H]⁺.

Example 3-4 (B) Preparation of4-isopropyl-6-{5-methanesulfonyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SFN-a02)

A methylene chloride (8 mL) solution of1,3-bis-benzyloxy-4-isopropyl-6-{5-methanesulfonyl-4-[4-(morpholin-4-ylmethyl)-phenyl]-4H-[1,2,4]triazol-3-yl}-benzene(1.05 g, 1.61 mmol: synthesized in the similar manner to that of theintermediate F45-05 of Example 2-2(B) was cooled to −20° C., mixed with1 N methylene chloride solution of boron trichloride (8 mL, 8.00 mmol)and warmed to room temperature in the course of 2 hours, and then thereaction was continued overnight. The reaction mixture was cooled againto −20° C., mixed with 1 N methylene chloride solution of borontrichloride (1 mL, 1.00 mmol) and warmed to room temperature in thecourse of 2 hours for the reaction to proceed. After completing thereaction, the reaction mixture was cooled to 0° C., and solid sodiumhydrogencarbonate was added until the pH was adjusted to about 7. Thereaction mixture was filtered to remove insoluble substances and washedwith a mixture (30 mL) of methylene chloride/methanol (10/1). Afterconcentrating under reduced pressure, the residue thus obtained waspurified by silica gel column chromatography (methylenechloride/methanol=10/1-2/1) to obtain the title compound (SFN-a02: 672mg, 88.1%) as white solids.

LC/MS (Method 1): m/z (ESI, POS): 473 [M+H]⁺; retention time: 2.19minutes.

¹H-NMR (400 MHz, CDCL₃, TMS) ppm: 7.61 (2H, d, J=8.4 Hz), 7.45 (2H, d,J=8.4 Hz), 6.53 (1H, s), 6.46 (1H, s), 3.74 (4H, brt, J=4.6 Hz), 3.61(2H, s), 3.50 (3H, s), 2.89 (1H, sept, J=6.8 Hz), 2.51 (4H, brt, J=4.6Hz), 0.74 (6H, t, J=7.0 Hz).

Example 3-5 Preparation of4-isopropyl-6-[5-methylsulfinyl-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFX-a08) and4-isopropyl-6-[5-methanesulfonyl-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN-a08)

4-isopropyl-6-[4-(4-methoxy-phenyl)-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diolwas produced according to Example 1-2 and Example 3-1 from5-isopropyl-2,4-bis-methoxymethoxybenzoic acid hydrazide [IM4-a (Example1-2)] in 4 steps.

4-isopropyl-6-[4-(4-methoxy-phenyl)-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(21.2 mg, 0.057 mmol) and methylene chloride (3 mL) were placed in atest tube and metachloroperbenzoic acid (29.5 mg, 0.171 mmol) was added,and the mixture was stirred at room temperature for 5 hours. Thereaction mixture was extracted with chloroform, and the organic layerwas washed with saturated sodium chloride solution and concentratedunder reduced pressure. The residue thus obtained was purified by HPLCfractionation to obtain the title compound (SFX-a08: 2.0 mg, 9.1%) andthe title compound (SFN-a08: 4.6 mg, 20.0%).

SFX-a08

LC/MS (Method 1): m/z (ESI, POS): 388 [M+H]⁺; retention time: 5.16minutes.

SFN-a08

LC/MS (Method 1): m/z (ESI, POS): 404 [M+H]⁺; retention time: 5.77minutes.

Example 3-7 Preparation of4-isopropyl-6-[4-(4-methoxy-phenyl)-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SMe-a08)

The title compound (SMe-a08: 23.7 mg, 39.4%) was obtained by a similarprocess to that of Example 3-9 using3-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-5-methylsulfanyl-4H-[1,2,4]triazole(Intermediate of Example 2-7, F60-01: 74.5 mg, 0.162 mmol) in place of5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-[4-(morpholin-4-ylmethyl)-phenyl]-2,4-dihydro-[1,2,4]triazol-3-one.

LC/MS (Method 3): m/z (ESI, NEG): 370 [M+H]⁺; retention time: 6.19minutes.

¹H-NMR (400 MHz, CD3CN, TMS) ppm: 7.19 (2H, d, J=9.0 Hz), 6.99 (2H, d,J=9.0 Hz), 6.73 (1H, s), 6.26 (1H, s), 3.82 (3H, s), 2.91 (1H, sept,J=7.0 Hz), 2.63 (3H, s), 0.94 (6H, d, J=7.0 Hz)

Example 3-8 Preparation of4-[5-(3-dimethylamino-ethylsulfanyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SR2-a08)

Preparation of{2-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-sulfanyl]-ethyl}-dimethyl-amine(F69-01): F69-01 was synthesized by a similar process to that of thefirst step of Example 3-16 using 2-dimethylamino-ethyl chloridehydrochloride in place of 3-dimethylamino-propyl chloride hydrochlorideof Example 3-16.

Preparation of4-[5-(3-dimethylamino-ethylsulfanyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SR2-a08)

The title compound (SR2-a08: 9.3 mg, 15.5%) was obtained by a similarprocess to that of Example 3-9 using F69-01 (72.3 mg, 0.054 mmol).

LC/MS (Method 3): m/z (ESI, POS): 429 [M+H]⁺; retention time: 3.66minutes.

Example 3-9 Preparation of4-isopropyl-6-(4-isopropyl-5-methanesulfonyl-4H-[1,2,4]triazol-3-yl)-benzene-1,3-diol(SFN-a21)

5 N hydrochloric acid (0.5 mL) was added to a methanol solution (0.5 mL)of4-isopropyl-3-(5-isopropyl-2,4-bis-methoxymethoxyphenyl)-5-methylsulfonyl-4H-[1,2,4]triazole(an intermediate of Example 2-13 F63-05: 35 mg, 82 μmol) and the mixturewas stirred for 8 hours. After cooling to ice cold conditions, it wasneutralized by adding saturated aqueous sodium hydrogencarbonate. Thesolution was extracted with ethyl acetate, and the extract was washedwith saturated sodium chloride solution, dried over anhydrous sodiumsulfate, and then sodium sulfate was removed by filtration and thesolvent was distilled off under reduced pressure. The solids thusobtained were dried under reduced pressure to obtain4-isopropyl-6-(4-isopropyl-5-methanesulfonyl-4H-[1,2,4]triazol-3-yl)-benzene-1,3-diol(SFN-a21, white solids, 20.8 mg, 75%).

LC/MS (Method 3): m/z (ESI, POS): 340 [M+H]⁺; retention time: 5.12minutes.

¹H-NMR [400 MHz, CDCl₃+CD₃OD (nine drops)] δ 1.19 (d, J=6.8 Hz, 6H),1.55 (d, J=7.0 Hz, 6H), 3.22 (sept., J=6.8 Hz, 1H), 3.60 (s, 3H), 4.80(sept., J=7.0 Hz, 1H), 6.38 (s, 1H), 7.04 (s, 1H).

Example 3-10 Preparation of4-isopropyl-6-[5-methanesulfonyl-4-(2-morpholin-4-ylethyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN-a26)

The bis(methoxymethyl) protected title compound (SFN-a26) was obtainedin 6 steps by a similar process to that of Example 2-2(B) using4-(2-aminoethyl)-morpholine in place of4-morpholin-4-ylmethyl-phenylamine (F45-000) of Example 2-2(B). Thiscompound was deprotected by a similar operation to that of Example 3-9to obtain the title compound (SFN-a26).

LC/MS (Method 3): m/z (ESI, POS): 428 [M+H]⁺; retention time: 6.56minutes.

¹H-NMR (400 MHz, CDCl₃) δ 1.25 (d, J=6.8 Hz, 6H), 2.40 (t, J=4.6 Hz,4H), 2.86 (t, J=6.6 Hz, 2H), 3.20 (sept., J=6.8 Hz, 1H), 3.58 (t, J=4.6Hz, 4H), 3.60 (s, 3H), 4.59 (t, J=6.6 Hz, 2H), 6.51 (s, 1H), 7.29 (s,1H).

Example 3-11 Preparation of4-isopropyl-6-[5-methanesulfonyl-4-[3-(morpholin-4-yl)-propyl]-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN-a27)

The intermediate, F93-05, of Example 2-16 was deprotected according toExample 3-9 to obtain the title compound (SFN-a27).

LC/MS (Method 4): m/z (ESI, POS): 425 [M+H]⁺; retention time: 3.80minutes.

¹H-NMR (400 MHz, CD₃OD, ppm): 7.09 (1H, s), 6.47 (1H, s), 4.37 (2H, m),3.54 (3H, s), 3.52 (4H, m), 3.20 (1H, m), 2.24 (2H, m), 2.16 (4H, brs),1.84 (2H, m), 1.19 (6H, d, J=6.96 Hz)

Example 3-125-(2,4-dihydroxy-5-isopropylphenyl)-4-[2-(1-oxopyridin-3-yl)ethyl]-3-methanesulfonyl-4H-[1,2,4]triazole(SFN-a29)

The bis(methoxymethyl) protected compound of the title compound(SFN-a29) was obtained in 6 steps by a similar process to that ofExample 2-2(B) using 3-(2-aminoethyl)pyridine in place of4-morpholin-4-ylmethyl-phenylamine (F45-000) of Example 2-2(B). Thiscompound was deprotected by a similar operation to that of Example 3-9to obtain the title compound (SFN-a29). LC/MS (Method 1): m/z (ESI,POS): 419 [M+H]⁺; retention time: 4.00 minutes.

¹H-NMR [400 MHz, CDCl₃—CD₃OD (three drops)] δ 1.19 (d, J=6.8 Hz, 6H),3.15-3.25 (m, 3H), 3.61 (s, 3H), 4.43-4.50 (m, 2H), 6.44 (s, 1H), 7.09(s, 1H), 7.25-7.27 (m, 2H), 7.94 (s, 1H), 8.06-8.10 (m, 1H).

Example 3-13 Preparation of1-{3-[3-(2,4-dihydroxy-5-isopropylphenyl)-5-methanesulfonyl[1,2,4]triazol-4-yl]-propyl}-pyrrolidine(SFN-a30)

N-(3-isothiocyanatopropyl)-2-pyrrolidinone was obtained by a similarprocess to that of Example 2-2(B) in 1 step usingN-(3-aminopropyl)-2-pyrrolidinone in place of4-morpholin-4-ylmethyl-phenylamine (F45-000) of Example 2-2(B). Usingthis compound, a bis(methoxymethyl) protected title compound (SFN-a30)was obtained in 5 steps by a similar process to that of Example 2-13.This compound was deprotected by a similar operation to that of Example3-9 to obtain the title compound (SFN-a30).

LC/MS (Method 1): m/z (ESI, POS): 423 [M+H]⁺; retention time: 4.73minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=5:1) b 1.20 (d, J=6.8 Hz, 6H), 1.90-2.00(m, 4H), 2.33 (t, J=8.1 Hz, 2H), 3.16-3.27 (m, 5H), 3.55 (s, 3H),4.19-4.26 (m, 2H), 6.42 (s, 1H), 7.12 (s, 1H).

Example 3-14 Preparation of4-isopropyl-6-[5-methanesulfonyl-4-(2-methoxy-ethyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN-a32)

A bis(methoxymethyl) protected title compound (SFN-a32) was obtained bya similar process to that of the fourth and fifth step of Example 2-12from the bis(methoxymethyl) protected compound (SH-a32) of Example 1-15.This compound was deprotected by a similar operation to that of Example3-9 to obtain the title compound (SFN-a32).

LC/MS (Method 3): m/z (ESI, POS): 356 [M+H]⁺; retention time: 4.69minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=3:1, ppm): 7.27 (1H, s), 6.44 (1s), 4.59(2H, t, J=5.49 Hz), 3.63 (2H, t, J=5.49 Hz), 3.54 (3H, s) 3.23 (1H, m),3.20 (3H, s), 1.20 (6H, d, J=6.77 Hz)

Example 3-15 Preparation of4-[4-(2-hydroxy-1-hydroxymethyl-ethyl)-5-methanesulfonyl-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SFN-a33)

The title compound (SFN-a33: white solids, yield 82%) was obtained bydeprotecting the intermediate F77-06 of Example 2-19 by a similarprocess to that of the seventh step of Example 2-19.

LC/MS (Method 1): m/z (ESI, POS): 372 [M+H]⁺, 394 [M+Na]⁺; retentiontime: 3.45 minutes.

¹H-NMR [400 MHz, DMSO-d₆, TMS] ppm: 1.115 (6H, d, J=7.0 Hz), 3.101 (1H,sept., J=7.0 Hz), 3.598 (3H, s), 3.674 (4H, bs), 4.58 (1H, b), 5.007(2H, bs), 6.480 (1H, s), 7.019 (1H, s), 9.705 (1H, s), 9.747 (1H, s).

Example 3-16 Preparation of4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SFN2-a08) hydrochloride

The First Step: Preparation of{3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-ylsulfanyl]-propyl}-dimethylamine(F68-01)

5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-2,4-dihydro-[1,2,4]triazol-3-thione(F53-04: 3.5 g, 7.9 mmol), potassium carbonate (2.6 g, 18.9 mmol) anddimethylformamide (150 mL) were placed in a 500 mL eggplant shapedflask, and then 3-dimethylaminopropyl chloride hydrochloride (1.49 g,9.4 mmol) was added, and the mixture was stirred at 100° C. for 2 hours.A similar operation was carried out using the same amount of materials,and after the reaction was completed, the mixture was cooled, mixed withsaturated sodium chloride solution (700 mL) and extracted twice withethyl acetate. The organic layers were combined, washed 4 times withsaturated sodium chloride solution (500 mL), dried with sodium sulfateand concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (methylene chloride/methanol) to obtainthe title compound (F68-01: 5.1 g, 61%).

LC/MS (Method 3): m/z (ESI, POS): 531 [M+H]⁺; retention time: 4.07minutes.

¹H-NMR (400 MHz, CDCl₃, TMS) ppm: 7.27 (1H, s), 7.06 (2H, d, J=8.8 Hz),6.86 (2H, d, J=8.8 Hz), 6.79 (1H, s), 5.15 (2H, s), 4.74 (2H, s), 3.79(3H, s), 3.46 (3H, s), 3.28 (2H, d, J=7.1 Hz), 3.22 (3H, s), 3.20 (1H,sept, J=7.0 Hz), 2.39 (2H, d, J=7.1 Hz), 2.22 (6H, s), 1.96 (2H, m),1.16 (6H, d, J=7.0 Hz)

The Second Step: Preparation of{3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazole-3-sulfanyl]-propyl}-dimethylamine(F68-02){3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-ylsulfanyl]-propyl}-dimethylamine(F68-01: 4.7 g, 8.9 mmol) and methylene chloride (350 mL) were placed ina 1000 mL eggplant shaped flask and cooled to 0° C., and a solution ofmetachloroperbenzoic acid (15.3 g, 88.7 mmol) in methylene chloride wasinstilled in 3 portions. After the reaction was completed, 10% aqueouspotassium bisulfite (100 mL) was added and the mixture was stirred for15 minutes. The organic layer was collected, washed twice with 1 Nsodium hydroxide (50 mL), dried with sodium sulfate and concentratedunder reduced pressure to obtain yellow solids. The solids thus obtainedwere subjected to the next reaction without purification.

LC/MS (Method 3): m/z (ESI, POS): 563 [M+H]⁺; retention time: 4.03minutes.

The Third Step: Preparation of4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SFN2-a08)

{3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazole-3-sulfonyl]-propyl}-dimethylamine(6.8 g, crude F68-02 in the previous step), ethanol (50 mL) and 3 Nhydrochloric acid (50 mL) were placed in a 200 mL eggplant shaped flaskand the mixture was stirred at 50° C. for 5 hours. After completing thereaction, the reaction mixture was neutralized with saturated sodiumhydrogencarbonate, extracted twice with ethyl acetate. The organic layerwas washed 4 times with saturated sodium chloride, dried with sodiumsulfate and then concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (methylenechloride/methanol), followed by basic silica gel column chromatography(methylene chloride/methanol) to obtain the title compound (F68-03: 1.5g, 35.7% in 2 steps)

LC/MS (Method 3): m/z (ESI, POS): 475 [M+H]⁺; retention time: 3.60minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 7.37 (2H, d, J=9.0 Hz), 7.00 (2H, d,J=9.0 Hz), 6.80 (1H, s), 6.30 (1H, s), 3.78 (3H, s), 3.45 (2H, d, J=7.7Hz), 2.96 (1H, sept, J=6.8 Hz), 2.27 (2H, d, J=7.5 Hz), 2.08 (6H, s),1.83 (2H, m), 0.94 (6H, d, J=6.8 Hz)

The Fourth Step: Preparation of4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SFN2-a08) hydrochloride

4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(700 mg, 1.5 mmol) and 1,4 dioxane (160 mL) were placed in a 300 mLeggplant shaped flask and the mixture was stirred at room temperature,and then 4 N hydrochloric acid/dioxane solution was slowly added andstirred for 20 minutes. After completing the reaction, depositedcrystals were collected by filtration, washed with hexane a few timesand dried under reduced pressure to obtain the title compound (SFN2-a08hydrochloride, 735 mg, 97.5%).

LC/MS (Method 3): m/z (ESI, POS): 475 [free M+H]⁺; retention time: 3.60minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 10.31 (1H, brs), 10.01 (1H, s), 9.83(1H, s), 6.80 (2H, d, J=9.0 Hz), 7.01 (2H, d, J=9.0 Hz), 6.80 (1H, s),6.35 (1H, s), 3.78 (3H, s), 3.69 (2H, d, J=7.5 Hz), 3.15 (2H, d, J=7.5Hz), 2.96 (1H, sept, J=6.8 Hz), 2.75 (6H, s), 2.16 (2H, m), 0.93 (6H, d,J=6.8 Hz),

IR(KBr): 2961, 1628, 1514, 1254, 1175, 1144, 623, 556.

Melting point: 233° C. (decomposition)

Example 3-17 Preparation of4-[5-(3-dimethylamino-propylsulfonyl)-4-(4-hydroxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SFN2-a11) trifluoroacetate

4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SNF2-a08 of Example 3-16: 47.5 mg, 0.1 mmol) and methylene chloride (5mL) were placed in a 10 mL eggplant-shaped flask, cooled to −78° C., andthen boron trichloride (0.4 mL) was added. The reaction mixture waswarmed slowly to room temperature in the course of 7 hours. Aftercompleting the reaction, methanol and sodium hydrogencarbonate wereadded, and inorganic substances were removed by filtration. The motherliquor was concentrated, and the residue thus obtained was purified byHPLC fractionation to obtain the title compound(SFN-a11•trifluoroacetate: 46.9 mg, 99%).

LC/MS (Method 3): m/z (ESI, POS): 461 [M+H]⁺; retention time: 3.02minutes.

Example 3-18 Preparation of4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(3-piperidin-1-yl-propane-1-sulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN3-a08) hydrochloride

The title compound (SFN3-a08 hydrochloride) can be obtained by a similarprocess to that of Example 3-16. That is, the title compound (SFN3-a08hydrochloride) was obtained in 4 steps from F53-04, by reacting1-(3-chloropropyl)piperidine hydrochloride, in place of3-dimethylaminopropyl chloride hydrochloride, to F53-04, and by carryingout the reactions sequentially as in Example 3-16.

LC/MS (Method 3): m/z (ESI, POS): 515 [M+H]⁺; retention time: 3.86minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=2:1, ppm): 7.34 (2H, d, J=8.97 Hz), 7.09(2H, d, J=8.98 Hz), 6.00 (1H, s), 6.40 (1H, s), 3.89 (3H, s), 3.68 (2H,d, J=7.14 Hz), 3.55 (2H, d, J=12.27 Hz), 3.28 (2H, m), 3.01 (1H, m),2.90 (2H, m), 2.41 (2H, m), 1.93 (5H, m), 1.51 (1H, m), 0.84 (6H, d,J=6.96 Hz)

Example 3-19 Preparation of4-(4-hydroxy-phenyl)-6-[4-isopropyl-5-(3-piperidin-1-yl-propane-1-sulfonyl)-4H-1,2,4-triazol-3-yl]-benzene-1,3-diol(SFN3-a11) trifluoroacetate

The title compound (SFN-all trifluoroacetate) was obtained by carryingout a similar reaction to that of Example 3-17 using4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(3-piperidin-1-yl-propane-1-sulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN3-a08) in place of4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SFN2-a08).

LC/MS (Method 3): m/z (ESI, POS): 501 [M+H]⁺; retention time: 3.33minutes.

Example 3-20 Preparation of4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(pyridin-3-ylmethanesulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN4-a08)

The title compound (SFN4-a08) can be obtained by a similar process tothat of Example 3-23. That is, the bis(methoxymethyl) protected titlecompound (SFN4-a08) and the bis(methoxymethyl) protected compound ofExample 3-21 (SFN5-a08) were obtained from F53-04 in 2 steps by reacting3-(bromomethyl)pyridine hydrobromide, in place of2-(bromomethyl)tetrahydro-2H-pyran, to F53-04, and carrying outsequentially the reactions similar to those of Example 3-23. Afterseparating both compounds, the bis(methoxymethyl) protected titlecompound (SFN4-a08) was deprotected by the similar operation to that ofExample 3-9 to obtain the title compound (SFN4-a08).

LC/MS (Method 3): m/z (ESI, POS): 481 [M+H]⁺; retention time: 4.39minutes.

¹H-NMR (400 MHz, CD₃OD, ppm): 8.68 (1H, d), 8.60 (1H, d, J=6.60 Hz),7.97 (1H, d, J=7.77 Hz), 7.77 (1H, m), 7.47 (2H, d, J=8.97 Hz), 7.16(2H, d, J=8.97 Hz), 6.76 (1H, s), 6.44 (1H, s), 4.98 (2H, m), 3.88 (3H,s), 3.00 (1H, m), 0.87 (3H, d, J=6.96 Hz), 0.86 (3H, d, J=6.96)

Example 3-21 Preparation of4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(1-oxy-pyridin-3-ylmethanesulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol (SFN5-a08)

The title compound (SFN5-a08) can be obtained by a similar process tothat of Example 3-23. That is, the bis(methoxymethyl) protected compoundof Example 3-21 (SFN4-a08) and the bis(methoxymethyl) protected titlecompound (SFN5-a08) were obtained from F53-04 in 2 steps by reacting3-(bromomethyl)pyridine hydrobromide, in place of2-(bromomethyl)tetrahydro-2H-pyran, to F53-04, and carrying outsequentially the reactions similar to those of Example 3-23. Afterseparating both compounds, the bis(methoxymethyl) protected titlecompound (SFN5-a08) was deprotected by the similar operation to that ofExample 3-9 to obtain the title compound (SFN5-a08).

LC/MS (Method 4): m/z (ESI, POS): 497 [M+H]⁺; retention time: 5.55minutes.

¹H-NMR (400 MHz, CD₃OD, ppm): 8.62 (1H, brs), 8.54 (1H, brs), 7.88 (1H,brs), 7.7 (1H, brs), 7.32 (2H, d, J=8.24 Hz), 7.05 (2H, d, J=8.24 Hz),6.71 (1H, s), 6.35 (1H, s), 5.13 (2H, m), 3.85 (3H, s), 3.00 (1H, m),0.88 (6H, d, J=6.77 Hz)

Example 3-22 Preparation of2-[5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-sulfonyl]-N,N-dimethyl-acetamide(SFN6-a08)

The First Step: Preparation of F81-02

Under an atmosphere of nitrogen, F53-04 (89 mg, 0.2 mmol), potassiumcarbonate (84 mg, 0.6 mmol), methyl bromoacetate (59 μL, 0.6 mmol) andmethanol (5 mL) were placed in a 50 mL flask, and the mixture was heatedfor 2 hours under reflux. The reaction mixture was mixed with water (30mL) and extracted twice with ethyl acetate (30 mL). The organic layerwas washed with saturated sodium chloride solution, dried with sodiumsulfate and concentrated under reduced pressure to obtain the crudetitle compound (F81-02: 101 mg).

LC/MS (Method 3): m/z (ESI, POS): 518 [M+H]⁺; retention time: 6.78minutes.

The Second Step: Preparation of F81-03

The crude preparation of F81-02 (101 mg), methanol (5 mL) and 1 Naqueous sodium hydroxide (0.25 mL) were placed in a 30 mL flask, and themixture was stirred at room temperature for 2 hours. Ethyl acetate (20mL) and 10% aqueous citric acid were added to the reaction mixture, andthe organic layer was separated. After washing with saturated sodiumchloride solution, the organic layer was dried with sodium sulfate andconcentrated under reduced pressure to obtain the crude title compound(F81-03, 100 mg).

LC/MS (Method 3): m/z (ESI, POS): 504 [M+H]⁺; retention time: 6.07minutes.

The Third Step: Preparation of F81-04

The crude preparation of F81-03 (100 mg) and dimethylformamide (3 mL)were placed in a 30 mL flask, and under ice cold conditions 50% aqueousdimethylamine (20 μL, 0.22 mmol), 1-hydroxybenzotriazole monohydrate (33mg, 0.24 mmol), and1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide•hydrochloride (46 mg,0.24 mmol) were sequentially added. After stirring overnight, thereaction mixture was mixed with water (30 mL) and extracted twice withethyl acetate (30 mL). The organic layer was washed sequentially with10% aqueous citric acid, saturated aqueous sodium hydrogencarbonate andsaturated sodium chloride, and then dried with sodium sulfate andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (chloroform:methanol=20:1) to obtain the titlecompound (F81-04: 76 mg, yield 71.6%, in total 3 steps).

LC/MS (Method 3): m/z (ESI, POS): 531 [M+H]⁺; retention time: 6.06minutes.

The Fourth Step: Preparation of F81-05

F81-04 (76 mg, 0.14 mmol) and dichloromethane (3 mL) were placed in a 30mL flask, and under ice cold conditions m-chloroperbenzoic acid (97 mg,0.56 mmol) was added, and the mixture was stirred for 20 hours.Chloroform (20 mL) and 10% aqueous potassium bisulfite (10 mL) wereadded to the reaction mixture, and after stirring the mixture for 10minutes, the organic layer was separated. The organic layer was washedwith saturated sodium chloride solution, dried with sodium sulfate andthen concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (chloroform:methanol=20:1) to obtainthe title compound (F81-05: 51 mg, yield 64.2%).

LC/MS (Method 3): m/z (ESI, POS): 563 [M+H]⁺; retention time: 6.11minutes.

The Fifth Step: Preparation of2-[5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-sulfonyl]-N,N-dimethyl-acetamide(SFN6-a08)

F81-05 (51 mg, 0.09 mmol) and methanol (3 mL) were placed in a 30 mLflask, and then 5 N hydrochloric acid (2 mL) was added, and the mixturewas stirred at 41° C. for 3 hours. The reaction mixture was concentratedand dissolved in methanol (3 mL), then mixed with silica gel (200 mg)concentrated under reduced pressure. The residue was purified by silicagel column chromatography (chloroform:methanol=20:1-10:1) to obtain thetitle compound (SFN6-a08: 25 mg, yield 58.5%).

LC/MS (Method 3): m/z (ESI, POS): 475 [M+H]⁺; retention time: 5.51minutes.

¹H-NMR (400 MHz, DMSO-d₆, ppm): 10.07 (1H, s), 9.79 (1H, s), 7.36 (2H,d, J=6.96 Hz), 7.01 (2H, d, J=6.96 Hz), 6.71 (1H, s), 6.32 (1H, s), 4.05(2H, s), 3.78 (3H, s), 2.96 (3H, s), 2.94 (1H, m), 2.81 (3H, s), 0.92(6H, d, J=7.2 Hz)

Example 3-23 Preparation of4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(tetrahydro-pyran-2-ylmethanesulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol (SFN7-a08)

The First Step Preparation of F82-02

F53-04 (67 mg, 0.15 mmol) and ethanol (4 mL) were placed in a 50 mLflask, and potassium carbonate (124 mg, 0.9 mmol) and2-(bromomethyl)tetrahydro-2H-pyran (58 μL, 0.45 mmol) were sequentiallyadded, and the mixture was heated for 4 hours under reflux. The reactionmixture was mixed with water (30 mL) and extracted twice with ethylacetate (30 mL). After washing with saturated sodium chloride, theorganic layer was dried with sodium sulfate and concentrated underreduced pressure to obtain the crude title compound (F82-02: 128 mg).

LC/MS (Method 3): m/z (ESI, POS): 544 [M+H]⁺; retention time: 7.14minutes.

The Second Step: Preparation of F82-03

After dissolving the crude F82-02 (128 mg) obtained in the first step indichloromethane (3 mL), m-chloroperbenzoic acid (104 mg, 0.60 mmol) wasadded, and the mixture was stirred for 20 hours. The reaction mixturewas mixed with chloroform (15 mL) and 10% aqueous potassium bisulfite(10 mL), and stirred for 10 minutes, and then the organic layer wasseparated. The organic layer was washed with saturated sodium chloridesolution, dried with sodium sulfate and then concentrated under reducedpressure. The residue was purified by silica gel column chromatography(chloroform:methanol=20:1) to obtain the title compound (F82-03: 72 mg,yield 83.3% in total 2 steps).

LC/MS (Method 3): m/z (ESI, POS): 576 [M+H]⁺; retention time: 6.90minutes.

The Third Step: Preparation of4-isopropyl-6-[4-(4-methoxy-phenyl)-5-(tetrahydro-pyran-2-ylmethanesulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol (SFN7-a08)

F82-03 (72 mg, 0.12 mmol) obtained in the second step was dissolved inmethanol (3 mL) and mixed with 5 N hydrochloric acid (2 mL), and themixture was stirred at 41° C. for 3 hours. After concentrating, thereaction mixture was dissolved in methanol (3 mL), mixed with silica gel(250 mg) and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (hexane:ethyl acetate=1:1)to obtain the title compound (SFN-a08: 38 mg, yield 64.9%).

LC/MS (Method 3): m/z (ESI, POS): 488 [M+H]⁺; retention time: 6.42minutes.

¹H-NMR (400 MHz, CDCl₃:CD₃OD=3:1, ppm): 7.41 (2H, d, J=8.98), 7.11 (2H,d, J=8.98), 6.47 (1H, s), 6.44 (1H, s), 6.71 (1H, s), 4.05 (4H, m), 3.91(3H, s), 3.71 (1H, m), 2.98 (1H, m), 1.86-1.42 (6H, m), 0.92 (6H, d,J=7.2 Hz)

Example 3-24 Preparation of4-isopropyl-6-{5-[2-(2-methoxy-ethoxy)-ethanesulfonyl]-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl}-benzene-1,3-diol(SFN8-a08)

The title compound (SFN8-a08) can be obtained by a similar process tothat of Example 3-23. That is, the title compound (SFN8-a08) wasobtained from F53-04 in 3 steps by reacting1-bromo-2-(2-methoxyethoxy)ethane, in place of2-(bromomethyl)tetrahydro-2H-pyran, to F53-04 and by carrying outsequentially the similar reactions as in Example 3-23.

LC/MS (Method 3): m/z (ESI, POS): 492 [M+H]⁺; retention time: 6.20minutes.

¹H-NMR (400 MHz, CDCl₃, ppm): 7.48 (2H, d, J=8.79 Hz), 7.18 (2H, d,J=8.79 Hz), 6.49 (1H, s), 6.47 (1H, s), 5.48 (1H, brs), 3.95 (2H, t,J=5.31 Hz), 3.89 (3H, s), 3.50 (4H, m), 3.26 (2H, m), 3.10 (3H, s), 2.91(1H, m), 0.79 (6H, d, J=6.8 Hz)

Example 3-25 Preparation of4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-isopropyl-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN2-a21) trifluoroacetate

The First Step: Preparation of{3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-isopropyl-4H-[1,2,4]triazol-3-ylsulfanyl]-propyl}-dimethylamine(F88-01)

5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-isopropyl-2,4-dihydro-[1,2,4]triazol-3-thione(F63-03: 114 mg, 0.3 mmol), potassium carbonate (249 mg, 1.8 mmol) andethanol (10 mL) were placed in a test tube, and next3-dimethylaminopropyl chloride hydrochloride (1.49 g, 9.4 mmol) wasadded. The mixture was stirred at 100° C. for 1 hour. After completingthe reaction, the reaction mixture was cooled naturally, mixed withsaturated sodium chloride solution and extracted twice with ethylacetate. The organic layers were combined, washed 4 times with saturatedsodium chloride solution, then dried with sodium sulfate andconcentrated under reduced pressure.

The residue thus obtained was subjected to the next reaction withoutpurification.

LC/MS (Method 3): m/z (ESI, POS): 467 [M+H]⁺; retention time: 4.09minutes.

The Second Step: Preparation of{3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-isopropyl-4H-[1,2,4]triazol-3-sulfanyl]-propyl}-dimethylamine(F88-02)

{3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-isopropyl-4H-[1,2,4]triazol-3-ylsulfanyl]-propyl}-dimethylamine(the crude product of the previous step (F88-01)) and methylene chloride(10 mL) were placed in a test tube, cooled to 0° C., and a solution ofmetachloroperbenzoic acid (0.62 g, 1.8 mmol) in methylene chloride wasinstilled in 3 portions. After completing the reaction, 10% aqueouspotassium bisulfite was added, and the mixture was stirred for 30minutes. After this, the organic layer was collected, washed twice with1 N sodium hydroxide, dried with sodium sulfate and concentrated underreduced pressure to obtain a liquid. The liquid thus obtained wassubjected to the next reaction without purification.

LC/MS (Method 6): m/z (ESI, POS): 499 [M+H]⁺; retention time: 6.86minutes.

The Third Step: Preparation of4-[5-(3-dimethylamino-propane-1-sulfonyl)-4-isopropyl-4H-1,2,4-triazol-3-yl]-6-isopropyl-benzene-1,3-diol(SFN2-a21) trifluoroacetate

{3-[5-(5-isopropyl-2,4-bis-methoxymethoxy-phenyl)-4-isopropyl-4H-1,2,4-triazol-3-sulfonyl]-propyl}-dimethylamine(6.8 g, unpurified in the previous step), ethanol (3 mL) and 5 Nhydrochloric acid (3 mL) were placed in a 200 mL eggplant shaped flaskand the mixture was stirred at room temperature for 10 hours. Aftercompleting the reaction, the reaction mixture was neutralized withsaturated sodium hydrogencarbonate and extracted twice with ethylacetate. The organic layer was washed 4 times with saturated sodiumchloride solution, dried with sodium sulfate and concentrated underreduced pressure. The residue was purified by HPLC fractionation toobtain the title compound (SFN2-a21•trifluoroacetate: 29 mg, 18.4%: in 3steps).

LC/MS (Method 4): m/z (ESI, POS): 411 [M+H]⁺; retention time: 4.22minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 9.85 (1H, brs), 9.55 (1H, brs), 6.97(1H, s), 6.52 (1H, s), 3.97 (2H, d, J=7.7 Hz), 3.32-3.22 (2H, m), 4.61(1H, sept, J=6.8 Hz), 3.12 (1H, sept, J=6.8 Hz), 2.29-2.20 (2H, m), 1.41(6H, d, J=6.8 Hz), 1.12 (6H, d, J=6.8 Hz)

Example 3-26 Preparation of4-isopropyl-6-[4-isopropyl-5-(3-piperidine-1-yl-propane-1-sulfonyl)-4H-[1,2,4]triazol-3-yl]-benzene-1,3-diol(SFN3-a21) trifluoroacetate

The title compound (SFN3-a21 trifluoroacetate) was obtained in 3 stepsby a similar process to that of Example 3-25 using1-(3-chloro-propyl)-piperazine hydrochloride in place of3-dimethylaminopropyl chloride hydrochloride.

LC/MS (Method 4): m/z (ESI, POS): 451 [M+H]⁺; retention time: 4.37minutes.

¹H-NMR (400 MHz, DMSO-d₆, TMS) ppm: 9.95 (1H, brs), 9.15 (1H, brs), 6.95(1H, s), 6.52 (1H, s), 4.61 (1H, sept, J=7.0 Hz), 3.97 (2H, d, J=7.9Hz), 3.52-3.43 (2H, m), 3.29-3.21 (2H, m), 3.21 (1H, sept, J=7.0 Hz),2.97-2.85 (2H, m), 2.36-2.24 (2H, m), 1.88-21.77 (2H, m), 1.68-1.55 (3H,m), 1.50-1.34 (2H, m), 1.41 (6H, d, J=7.0 Hz), 1.12 (6H, d, J=7.0 Hz)

Example 3-27 Preparation ofN-[5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-methanesulfonamide(N1-a08)

The First Step: Preparation of F62-02 F62-01 (synthesized in a similarmanner to the intermediate F63-05 of Example 2-3: 362 mg, 0.62 mmol),methane sulfonamide (117 mg, 1.86 mmol), potassium carbonate (514 mg,3.72 mmol) and dimethylsulfoxide (3.5 mL) were placed in a 30 mL flaskand the mixture was stirred at 90° C. for 120 hours. The reactionmixture was mixed with water (50 mL), and after adjusting the pH to 7.5with 2 N hydrochloric acid, the mixture was extracted twice with ethylacetate (100 mL). The organic layer was washed with saturated sodiumchloride solution, dried with sodium sulfate and then concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (hexane:ethyl acetate=1:1-1:2) to obtain the titlecompound (F62-02: 108 mg, yield 29.0%).

LC/MS (Method 5): m/z (ESI, POS): 599 [M+H]⁺; retention time: 6.84minutes.

The Second Step: Preparation ofN-[5-(2,4-dihydroxy-5-isopropyl-phenyl)-4-(4-methoxy-phenyl)-4H-[1,2,4]triazol-3-yl]-methanesulfonamide(N1-a08)

After dissolving F62-02 (99 mg, 0.165 mmol) in anhydrous dichloromethane(3 mL), the solution was cooled to −20° C. and mixed with 1 moltrichloroborane solution (3 mL). After warming to 0° C., the mixture wasstirred for 2 hours. The reaction mixture was mixed with methanol (3 mL)and then with solid sodium hydrogencarbonate until pH became about 7.0with pH paper. The reaction mixture was filtered and the insolublefraction was washed sufficiently with chloroform:methanol (3:1). Thefiltrate and the wash were combined and concentrated, and the residuewas purified by silica gel column chromatography(chloroform:methanol=15:1-10:1) to obtain the title compound (N1-a08:44.2 mg, yield 63.9%).

LC/MS (Method 3): m/z (ESI, POS): 419 [M+H]⁺; retention time: 4.93minutes.

¹H-NMR (400 MHz, DMSO-d₆, ppm): 13.04 (1H, brs), 9.44 (1H, brs), 7.15(2H, d, J=8.98 Hz), 6.92 (2H, d, J=8.98), 6.89 (1H, s), 6.24 (1H, s),4.04 (H, s), 3.73 (3H, s), 2.98 (1H, m), 2.87 (3H, s), 1.00 (3H, d,J=6.96)

Test Example 1

To confirm the binding of the compound of the present invention toHSP90, an HSP90 binding assay system was constructed using BIACORE (adevice for measuring biomolecular binding activity which measures thebinding of biomolecules in real time by reproducing the biomolecularbinding on a sensor chip applying Surface Plasmon Resonance (SPR))(refer to Adamczyk, M., Moore, J. A., Yu, Z. (2000) Methods, 20, p.319-328).

The HSP90 binding assay system is a system for detecting the change ofmass as SPR signal caused by the binding between herbimycin A fixed on asensor chip and rHSP90 by BIACORE-X, in which 17-(6-aminohexylamino)herbimycin A is fixed through a carboxyl group on the surfaceof the sensor chip (CM5, BIACORE) on which carboxymethyldextran has beenintroduced. The method was according to the protocol of BIACORE. Inaddition, 17-(6-aminohexylamino)herbimycin A was synthesizes asdescribed below.

Reagent Synthesis Example 1 Preparation of17-(6-aminohexylamino)herbimycin A

17-(6-aminohexylamino)herbimycin A (17(A)) was synthesized according tothe following scheme (4).

Herbimycin A (472 mg, 0.82 mmol) was dissolved in chloroform (42 mL),mixed with hexamethylenediamine (691 mg, 11.9 mmol) and the mixture wasstirred at room temperature for 18 hours. Water (50 mL) and chloroform(50 mL) were added to the reaction mixture, and the chloroform layer wasseparated. The chloroform layer was washed with saturated sodiumchloride solution, dried with sodium sulfate, and the solvent wasdistilled off. The residue was subjected to silica gel columnchromatography (150 mL, chloroform:methanol: acetic acid=30:6:1) toobtain the crude compound (80 mg). Further, NH-silica gel columnchromatography (40 mL, chloroform, Fuji Sylisia Chemical Ltd.) wasperformed to obtain 17-(6-aminohexylamino)herbimycin A (17(A)) (48 mg,yield 8.5%).

LC/MS: m/z (ESI,POS): 689 [M+H]⁺

¹H-NMR (200 MHz, CDCl₃) ppm: 9.52 (1H, br), 7.70 (1H, br), 7.00 (1H, s,H-19), 7.00 (1H, d, J=11.3 Hz), 6.51 (1H, t, J=10.9 Hz, H-4), 5.84 (1H,dd, J=10.9 and 7.3 Hz, H-5), 5.4-5.6 (2H, br, H-7 and H9), 4.75 (2H, br,CONH₂), 4.71 (1H, s, H-15), 4.48 (1H, d, J=7.3 Hz, H-6), 3.68 (2H, m),3.52 (3H, OMe), 3.35 (3H, OMe), 3.33 (3H, OMe), 3.32 (3H, OMe), 2.00(3H, s), 1.95-1.22 (11H, m), 1.68 (3H, s, Me), 1.07 (3H, d, J=6.8 Hz),0.96 (3H, d, J=6.5 Hz)

rHSP90 (Stressgen Biotchnologies Corp., Victoria, BC Canada) at 50 μg/mLwas exposed for 10 seconds to the surface of the sensor chip on whichherbimycin A was fixed and the SPR signal (numerical value ofinteraction) was detected. As the result, the increase of the SPR signalwas observed, confirming the binding of rHSP90 and herbimycin A fixed onthe sensor chip.

After mixing rHSP90 protein (5×10⁻⁷M (50 μg/mL)) with the triazolederivative of the present invention, the mixture was exposed for 10seconds to the surface of the sensor chip on which herbimycin A wasfixed, and the SPR signal was measured by BIACORE-X.

Next, using following formula (1), the inhibitory activity (bindinginhibition rate (%)) of the compound of the present invention to thebinding between rHSP90 and fixed herbimycin A was obtained.

binding inhibition rate (%)=((b−s)/b)×100  Formula (1)

wherein b is the SPR signal when the compound of the present inventionis not added to the sample: s is the SPR signal when the compound of thepresent invention is added to the sample.

From the concentration of the triazole derivative of the presentinvention and its inhibitory activity, the concentration, at which thebinding between HSP90 protein and fixed herbimycin A was inhibited by50%, was calculated and termed an IC50 value.

The triazole derivatives of the present invention reduced the SPRsignal, concentration dependently, indicating that the compounds of thepresent invention inhibited the binding between HSP90 and fixedherbimycin A. Table 4-1-Table 6-4 show IC50 values of the bindinginhibition.

Comparative Example 1

As comparative examples, the inhibitory activity (binding inhibitionrate (%)) against the binding between rHSP90 and fixed herbimycin A ofSH-b04, SH-b05 and SH-b06 (all from Scientific Exchange Co.), which areshown below, were measured in a similar manner to the triazolederivatives of the present invention. Table 7 shows IC 50 values of thebinding inhibition.

Test Example 2 Measurement Test for Amount of HSP90 Client Protein

MCF7 cells (American Type Culture Collection, Rockville, Md.) weretreated with the compounds of the present invention at variousconcentrations for 16 hours to confirm that the HSP90 inhibitors of thepresent invention induce a depression of intracellular concentration ofthe client protein or target polypeptide that bind to HSP90. The amountof the HSP90 client protein Her2 and ERα was evaluated by the Westernblotting method.

One million cells were seeded in a 6 cm dish, and 24 hours later thecompounds of the present invention (Example 1-2, Example 2-1, Example2-13, Example 2-2, Example 3-4, Example 2-5, Example 3-16, Example 3-6,Example 2-7, Example 3-27, Example 2-10 and Example 3-18) were added.Cells treated with the compounds were washed, mixed with 150 μl of lysisbuffer (RIPA, 150 mM NaCl, 1% NP40, 0.1% deoxycholate (sodium salt),0.1% SDS, 1 mM EDTA, 10 mM Tris-HCl (pH 8.0)) and incubated at 4° C. for30 minutes. The cell lysates were centrifuged (15,000 rpm, 20 minutes)and protein (20 μg) of the supernatant was subjected to SDSpolyacrylamide gel electrophoresis (SDS-PAGE). After finishing theelectrophoresis, proteins in the gel were transferred to a PVDFmembrane. The membrane after the transfer was treated with a primaryantibody against the HSP90 client protein (anti Her2 or anti ERαantibody, both from Santa Cruz Biotechnology, Santa Cruz, Calif.), thentreated with secondary antibodies (anti-rabbit Ig, horse radishperoxidase conjugated F(ab′)2 fragment (derived from donkey): AmershamBiosciences, UK Limited, Buckinghamhshire, UK.) and then the amount ofthe client protein was detected as the intensity of chemiluminescencesignal with a chemiluminescence reagent (ECL: Amersham Biosciences UKLimited, Buckinghamshire, UK). FIG. 1-FIG. 7 shows the results ofelectrophoresis.

Test Example 3

Human breast cancer cells (MCF7) were treated with samples of variousconcentrations of known HSP90 inhibitors (that is, geldanamycin,herbimycin, radicicol and PU3) and the compounds of the presentinvention for 72 hours to confirm the effect of the triazole derivativesof the present invention on the cell growth. The cell ratio after thedrug treatment was evaluated by measuring 660 nm absorbance by amicroplate reader (BioRad) after staining the cells with the methyleneblue method.

Cells were distributed to wells of 96 well plates at 2000 cells per welland treated with drugs 24 hours later. Further, the medium was removed72 hours later and the cells were fixed by adding 50 μL of methanol andstanding at room temperature for 2 minutes. After removing methanol, 100μL of a staining solution was added and stained for 30 minutes. Afterwashing 3 times with 200 μL of distilled water, 3% HCl solution wasadded, and the 660 nm absorbance of methylene blue was measured by amicroplate reader (BioRad).

Next, the antiproliferative activity rate (%) was obtained by theformula (2) below.

Antiproliferative activity rate (%)=((B−A)/B)×100  Formula (2)

wherein B is 660 nm absorbance of the sample when the compound of thepresent invention is not added, and A is 660 nm absorbance of the samplewhen the compound of the present invention is added.

From the concentration of the triazole derivative of the presentinvention and the antiproliferative activity rate, the concentration wasobtained using the above formula, at which the cell growth wassuppressed by 50% by comparing with the control, and termed an IC50value.

The IC50 value of the HSP90 inhibitors, geldanamycin, herbimycin,radicicol and PU3 was 0.012 μM, 0.16 μM, 0.019 μM and 91 μM,respectively. The IC 50 values for antiproliferative activity of thetriazole derivatives of the present invention are shown in Tables4-1-6-4.

Comparative Example 2

The antiproliferative activity rate (%) was obtained for SH-b04, SH-b05and SH-b06 described above by a similar manner to that of the triazolederivatives of the present invention. Table 7 shows IC 50 values ofantiproliferative activity.

The above results have made it clear that the triazole derivatives ofthe present invention have a antiproliferative activity on MCF7 cellsand that their antiproliferative effect is superior compared to that ofother compounds known to have the HSP90 inhibitory activity.

The above results have made it clear that the triazole derivatives ofthe present invention have the HSP90 inhibitory activity as well as theantiproliferative activity against cancer cells, and are useful astherapeutic drugs for cancer.

Test Example 4 Antitumor Effect on Nude Mice Transplanted with HumanLung Cancer

Human lung cancer H460 tumor fragment, which had been passagedsubcutaneously in nude mice, was made into about 3 mm fragment andtransplanted subcutaneously to the back of nude mice using a trocar.When the tumor volume grew to about 50 mm³ or above, the compound of thepresent invention was administered to the tail vein once a day for 5 or7 days every day. The compound of the present invention was dissolved inDMSO, mixed with TWEEN 80, and then diluted with a 5% glucose injectionsolution and used. The vehicle of the compound was administered to thecontrol group of OH-a01. The control groups in other experiments werenot treated. The tumor volume was measured on the first administrationday and on the evaluation day (7 or 8 days after the start of theadministration), and the relative tumor volume of the evaluation day wasobtained from the tumor volume on the first administration day. Further,the tumor volume was calculated by measuring the major axis (Lmm) andminor axis (Wmm) of the tumor and by the formula (L×W²)/2. The resultsare shown in Tables 8, 9 and 10.

Table 8 shows the anti-tumor effect (7 days continuous intravenousadministration) of OH-a01 (Example 2-1) on human lung cancer H460.Relative tumor volume represents the average relative tumor volume onday 7 after the start of the administration, assuming the tumor volumeof the starting day is 1.0.

Table 9 shows the anti-tumor effect (5 days continuous intravenousadministration) of OH-a02 (Example 2-2) and OH-a08 (Example 2-7) onhuman lung cancer H460. Relative tumor volume represents the averagerelative tumor volume on day 8 after the start of the administration,assuming the tumor volume of the starting day is 1.0.

Table 10 shows the anti-tumor effect (5 days continuous intravenousadministration) of OH-a13 (Example 2-5) and SFN3-a08 (Examples 3-18) onhuman lung cancer H460. Relative tumor volume represents the averagerelative tumor volume on day 8 after the start of the administration,assuming the tumor volume of the starting day is 1.0.

INDUSTRIAL APPLICABILITY

It was made it clear that the compounds of the present inventionconcentration dependently inhibited the tumor growth of human lungcancer H460 transplanted on nude mice and that they are useful asanti-cancer agent.

TABLE 1 Compounds represented by General formula (1) or General formula(4)

SH-a01 Example 1-1

SH-a01 Example 1-1

SH-a02 Example 1-2

SH-a03 Example 1-3

SH-c02 Example 1-4

SH-d01 Example 1-5

SH-a08 Example 1-6

SH-a15 Example 1-7

SH-a16 Example 1-8

SH-a21 Example 1-9

SH-a22 Example 1-10

SH-a23 Example 1-11

SH-a25 Example 1-12

SH-a28 Example 1-13

SH-a31 Example 1-14

SH-a32 Example 1-15

SH-f08 Example 1-16

TABLE 2 Compounds represented by General formula (1) or General formula(4)

OH-a01 Example 2-1

OH-a02 Example 2-2

OH-c02 Example 2-3

OH-e02 Example 2-4

OH-a13 Example 2-5

OH-a14 Example 2-6

OH-a08 Example 2-7

OH-a09 Example 2-8

OH-a10 Example 2-9

OH-a11 Example 2-10

OH-a12 Example 2-11

OH-a17 Example 2-12

OH-a21 Example 2-13

OH-a24 Example 2-14

OH-a26 Example 2-15

OH-a27 Example 2-16

OH-a30 Example 2-17

OH-a32 Example 2-18

OH-a33 Example 2-19

TABLE 3-1 Compounds represented by General formula (1) or Generalformula (4)

SMe-a02 Example 3-1

SFX-a07 Example 3-2

SFN-a07 Example 3-2

SMe-d01 Example 3-3

SFN-a02 Example 3-4

SFX-a08 Example 3-5

SFN-a08 Example 3-6

SMe-a08 Example 3-7

SR2-a08 Example 3-8

SFN-a21 Example 3-9

SFN-a26 Example 3-10

SFN-a27 Example 3-11

SFN-a29 Example 3-12

SFN-a30 Example 3-13

SFN-a32 Example 3-14

SFN-a33 Example 3-15

SFN2-a08 Example 3-16

SFN2-a11 Example 3-17

SFN3-a08 Example 3-18

SFN3-a11 Example 3-19

TABLE 3-2 Compounds represented by General formula (1) or Generalformula (4)

SFN4-a08 Example 3-20

SFN5-a08 Example 3-21

SFN6-a08 Example 3-22

SFN7-a08 Example 3-23

SFN8-a08 Example 3-24

SFN2-a21 Example 3-25

SFN3-a21 Example 3-26

N1-a08 Example 3-27

TABLE 4-1 HSP90 inhibitory activity and antiproliferative activity (1)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SH-a01-TF Example 1-1

0.19 0.020 SH-a01 Example 1-1

0.14 0.014 SH-a02 Example 1-2

0.24 0.036 SH-a03 Example 1-3

0.15 0.15  SH-c02-TF Example 1-4

0.25 0.48  SH-d01 Example 1-5

0.15 0.58  SH-a08 Example 1-6

 0.085 0.057 SH-a15 Example 1-7

0.25 0.040

TABLE 4-2 HSP90 inhibitory activity and antiproliferative activity (1)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SH-a16 Example 1-8

0.14 0.24 SH-a21 Example 1-9

0.22 0.40 SH-a22 Example 1-10

0.27 0.45 SH-a23 Example 1-11

0.42 0.33 SH-a25•TFA Example 1-12

1.0  0.89 SH-a28 Example 1-13

0.43 7.2  SH-a31 Example 1-14

0.31 0.55 SH-a32 Example 1-15

0.24 0.55

TABLE 4-3 HSP90 inhibitory activity and antiproliferative activity (1)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SH-f08 Example 1-16

0.56 3.0

TABLE 5-1 HSP90 inhibitory activity and antiproliferative activity (2)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity OH-a01 Example 2-1

0.14  0.0049 OH-a02 Example 2-2

0.18 0.016 OH-c02 Example 2-3

0.22 0.14  OH-a02 Example 2-4

0.15 0.24  OH-a13•TFA Example 2-5

0.25 0.071 OH-a14 Example 2-6

0.19 0.72  OH-a08 Example 2-7

0.12 0.011

TABLE 5-2 HSP90 inhibitory activity and antiproliferative activity (2)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity OH-a09 Example 2-8

0.16  0.0067 OH-a10 Example 2-9

0.21 0.014 OH-a11 Example 2-10

0.22 0.077 OH-a12 Example 2-11

0.19 >20     OH-a17 Example 2-12

0.22 0.026 OH-a21 Example 2-13

0.13 0.038 OH-a24•TFA Example 2-14

1.1  0.79  OH-a26•HCl Example 2-15

0.18 0.60 

TABLE 5-3 HSP90 inhibitory activity and antiproliferative activity (2)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity OH-a27 Example 2-16

0.24 0.22 OH-a30 Example 2-17

0.14 1.1  OH-a32 Example 2-18

0.35 0.39 OH-a33 Example 2-19

0.37 3.3 

TABLE 6-1 HSP90 inhibitory activity and antiproliferative activity (3)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SMe-a02-TF Example 3-1

0.47 0.53 SFX-a07-TF Example 3-2

0.064 12    SFN-a07 Example 3-2

0.32 24    SMe-d01-TF Example 3-3

0.60 5.5  SFN-a02 Example 3-4

0.12 0.15 SFX-a08 Example 3-5

0.12 0.36 SFN-a08 Example 3-5

 0.094 0.27

TABLE 6-2 HSP90 inhibitory activity and antiproliferative activity (3)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SMe-a08 Example 3-7

 0.59 3.1 SR2-a08 Example 3-8

 0.74 1.8 SFN-a21 Example 3-9

 0.23 16   SFN-a26 Example 3-10

3.4 >20    SFN-a27 Example 3-11

1.6 >20    SFN-a29 Example 3-12

3.6 >20    SFN-a30 Example 3-13

2.4 >20   

TABLE 6-3 HSP90 inhibitory activity and antiproliferative activity (3)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SFN-a32 Example 3-14

1.9  >20     SFN-a33 Example 3-15

1.3  15    SFN2-a08•TFA Example 3-16

0.70 0.069 SFN2-a11•TFA Example 3-17

0.43 0.098 SFN3-a08 Example 3-18

0.59 0.044 SFN3-a11•TFA Example 3-19

0.73 0.044 SFN4-a08 Example 3-20

0.23 9.9 

TABLE 6-4 HSP90 inhibitory activity and antiproliferative activity (3)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SFN5-a08 Example 3-21

0.43 3.9 SFN6-a08 Example 3-22

0.58  0.55 SFN7-a08 Example 3-23

0.72  0.72 SFN8-a08 Example 3-24

0.60 2.4 SFN2-a21•TFA Example 3-25

1.0  5.2 SFN3-a21•TFA Example 3-26

1.4  4.1 N1-a08 Example 3-27

0.29  0.18

TABLE 7 HSP90 inhibitory activity and antiproliferative activity (4)IC50 (μM) Test Example 1 HSP90 Test Example 2 inhibitoryAntiproliferative No. Structure activity activity SH-b04 ComparativeExample (1)

0.17 5.7 SH-b05 Comparative Example (2)

0.20 9.0 SH-b06 Comparative Example (3)

0.69 77  

TABLE 8 Administration dose Relative tumor volume Compound (mg/kg/day)(Average ± SD) Control group 0 7.3 ± 1.7 Example 2-1 7.5 1.7 ± 0.5 3.83.3 ± 0.7

TABLE 9 Administration dose Relative tumor volume Compound (mg/kg/day)(Average ± SD) Control group 0 4.6 ± 3.1 Example 2-2 150 2.1 ± 0.1 753.5 ± 2.3 Example 2-7 200 1.1 ± 0.3 100 1.9 ± 1.0

TABLE 10 Administration dose Relative tumor volume Compound (mg/kg/day)(Average ± SD) Control group 0 6.5 ± 2.0 Example 2-5 100 1.1 ± 0.3 501.4 ± 0.5 Example 3-18 150 2.9 ± 0.6 100 5.4 ± 1.3

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a depression of the amount of HSP90 client protein Her2 andERα by the HSP90 inhibitor of the present invention (Example 1-2 andExample 2-1) in the protein assay for the HSP90 client protein in Testexample 2. After treating MCF7 cells with 4 or 5 concentrations of eachHSP90 inhibitor for 16 hours, the amount of each Her2 and ERα protein inthe lysate obtained from the cells was evaluated by Western blottingmethod. β-actin is the intra-cellular standard substance. Cont.represents a control;

FIG. 2 shows a depression of the amount of HSP90 client protein Her2 andERα by the HSP90 inhibitor of the present invention (Example 2-2 andExample 2-13) in the protein assay for the HSP90 client protein in Testexample 2. After treating MCF7 cells with 4 concentrations of each HSP90inhibitor for 16 hours, the amount of each Her2 and ERα protein in thelysate obtained from the cells was evaluated by Western blotting method.β-actin is the intra-cellular standard substance. Cont. represents acontrol;

FIG. 3 shows a depression of the amount of HSP90 client protein Her2 andERα by the HSP90 inhibitor of the present invention (Example 3-4) in theprotein assay for the HSP90 client protein in Test example 2. Aftertreating MCF7 cells with 4 concentrations of each HSP90 inhibitor for 16hours, the amount of each Her2 and ERα protein in the lysate obtainedfrom the cells was evaluated by Western blotting method. β-actin is theintra-cellular standard substance. Cont. represents a control;

FIG. 4 shows a depression of the amount of HSP90 client protein Her2 andERα by the HSP90 inhibitor of the present invention (Example 2-5 andExample 3-16) in the protein assay for the HSP90 client protein in Testexample 2. After treating MCF7 cells with 4 concentrations of each HSP90inhibitor for 16 hours, the amount of each Her2 and ERα protein in thelysate obtained from the cells was evaluated by Western blotting method.β-actin is the intra-cellular standard substance. Cont. represents acontrol;

FIG. 5 shows a depression of the amount of HSP90 client protein Her2 andERα by the HSP90 inhibitor of the present invention (Example 3-6) in theprotein assay for the HSP90 client protein in Test example 2. Aftertreating MCF7 cells with 3 concentrations of each HSP90 inhibitor for 16hours, the amount of each Her2 and ERα protein in the lysate obtainedfrom the cells was evaluated by Western blotting method. β-actin is theintra-cellular standard substance. Cont. represents a control;

FIG. 6 shows a depression of the amount of HSP90 client protein Her2 andERα by the HSP90 inhibitor of the present invention (Example 2-7 andExample 3-27) in the protein assay for the HSP90 client protein in Testexample 2. After treating MCF7 cells with 4 concentrations of each HSP90inhibitor for 16 hours, the amount of each Her2 and ERα protein in thelysate obtained from the cells was evaluated by Western blotting method.β-actin is the intra-cellular standard substance. Cont. represents acontrol; and

FIG. 7 shows a depression of the amount of HSP90 client protein Her2 andERα by the HSP90 inhibitor of the present invention (Example 2-10 andExample 3-18) in the protein assay for the HSP90 client protein in Testexample 2. After treating MCF7 cells with 3 concentrations of each HSP90inhibitor for 16 hours, the amount of each Her2 and ERα protein in thelysate obtained from the cells was evaluated by Western blotting method.β-actin is the intra-cellular standard substance. Cont. represents acontrol.

1. A compound represented by the following structural formula (1):

or a pharmaceutically acceptable salt, or a prodrug thereof, wherein: Ris an optionally substituted heterocyclic aryl or an optionallysubstituted carbocyclic aryl group; R₁ and R₂ are —H, an acyl group, acarbamoyl group, an alkoxycarbonyl group, or an alkoxymethyl group; X isan optionally substituted alkyl group, an optionally substituted alkenylgroup, an optionally substituted alkynyl group, cyano group, halogenatom, nitro group, an optionally substituted cyclic alkyl group, anoptionally substituted carbocyclic aryl group, an optionally substitutedheterocyclic aryl group, hydroxyl group, an optionally substitutedalkoxyl group, an optionally substituted aryloxy group, mercapto group,an optionally substituted alkylthio group, an optionally substitutedarylthio group, an optionally substituted amino group, an optionallysubstituted carbamoyloxy group, an optionally substituted ureido group,an optionally substituted acyloxy group, an optionally substitutedacylamino group, an optionally substituted alkoxycarbonyloxy group, anoptionally substituted alkoxycarbonylamino group, an optionallysubstituted sulfonylamino group, an optionally substitutedsulfamoylamino group, an optionally substituted acyl group, anoptionally substituted carboxyl group, an optionally substitutedalkoxycarbonyl group, an optionally substituted carbamoyl group, anoptionally substituted sulfamoyl group, an optionally substitutedalkylsulfonyl group, an optionally substituted arylsulfonyl group, anoptionally substituted alkylsulfinyl group, or an optionally substitutedarylsulfinyl group; and Y is —OH, —SH, an optionally substituted acyloxygroup, an optionally substituted alkoxycarbonyloxy group, or anoptionally substituted carbamoyloxy group; provided that when Y is —OHor —SH at least one of R₁ and R₂ is not hydrogen.
 2. The compound ofclaim 1, wherein the compound is not3-(2,4-Dihydroxy-5-ethyl-phenyl)-4-(3-trifluoromethyl-phenyl)-5-carbamoyloxy-[1,2,4]triazole;3-(2,4-Dihydroxy-5-ethyl-phenyl)-4-(1-methyl-indol-4-yl)-5-carbamoyloxy-[1,2,4]triazole;3-(2,4-Dihydroxy-5-methoxy-phenyl)-4-(8-methoxy-quinolin-5-yl)-5-carbamoyloxy-[1,2,4]triazole;3-(2-Hydroxy-4-ethoxycarbonyoxy-5-methoxy-phenyl)-4-(1-isopropyl-benzoimidazol-4-yl)-5-hydroxy-[1,2,4]triazole;3-(2-Hydroxy-4-ethoxycarbonyoxy-5-ethyl-phenyl)-4-(naphthalin-2-yl)-5-hydroxy-[1,2,4]triazole;3-[2-Hydroxy-4-(dimethyl-carbamoyoxy)-5-ethyl-phenyl]-4-(naphthalin-2-yl)-5-hydroxy-[1,2,4]triazole;3-[2-Hydroxy-4-(dimethyl-carbamoyoxy)-5-chloro-phenyl]-4-(quinolin-5-yl)-5-mercapto-[1,2,4]triazole;3-[2-Hydroxy-4-(dimethyl-carbamoyoxy)-5-ethyl-phenyl]-4-(2,3-difluoro-phenyl)-5-mercapto-[1,2,4]triazole;or3-[2-Hydroxy-4-isobutyryloxy-5-ethyl-phenyl]-4-(1-methyl-benzo-imidazol-4-yl)-5-hydroxy-[1,2,4]triazole.3. The compound of claim 1, wherein when Y is —OH or —SH, R₁ and R₂ areselected from the group consisting of an acyl group, a carbamoyl group,an alkoxycarbonyl group, and an alkoxymethyl group.
 4. The compound ofclaim 1, wherein R₁ and R₂ are the same.
 5. The compound of claim 1,wherein R is an optionally substituted carbocyclic aryl group.
 6. Thecompound of claim 1, wherein R is an optionally substituted heterocyclicaryl group.
 7. The compound of claim 1, wherein R has the followinggeneral formula:

m is an integer from 0 to 5; and A is an optionally substituted cyclicor non-cyclic amino group, an optionally substituted cyclic ornon-cyclic acylamino group, or an optionally substituted cyclic ornon-cyclic sulfonylamino group.
 8. The compound of claim 1, wherein Rhas the following general formula:

m is an integer from 0 to 5; and A is an optionally substituted cyclicor non-cyclic amino group, an acylamino group, or a sulfonylamino group.9. The compound of claim 1, wherein X is an optionally substituted C₁₋₈alkyl group, an optionally substituted C₁₋₈ alkoxyl group, or anoptionally substituted C₁₋₈ alkylthio group.
 10. The compound of claim1, wherein Y is an acyloxy group, an alkoxycarbonyloxy group, or acarbamoyloxy group.
 11. A pharmaceutical composition comprising: apharmaceutically acceptable carrier and a compound according to claim 1.12. A compound represented by the following structural formula (I):

or a pharmaceutically acceptable salt, or a prodrug thereof, wherein: Ris an optionally substituted heteroaryl or an optionally substitutedaryl; X is an optionally substituted alkyl, an optionally substitutedalkenyl, an optionally substituted alkynyl, cyano, halo, nitro, anoptionally substituted cycloalkyl, haloalkyl, optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted aralkyl, an optionally substitutedheteroaralkyl, —OR₇, —SR₇, —NR₁₀R₁₁, —OC(O)NR₁₀ ^(a)R₁₁ ^(a),—NR₇C(O)NR₁₀ ^(a)R₁₁ ^(a), OC(O)R₇ ^(a), NR₇ ^(b)C(O)R₇ ^(b′), —OC(O)OR₇^(c), —NR₇ ^(b)C(O)OR₇ ^(b′), —OCH₂C(O)NH₂, SCH₂C(O)NH₂, NR₇ ^(b)S(O)₂R₇^(b′), —NR₇ ^(b)S(O)₂NR₁₀ ^(a)R₁₁ ^(a), —C(O)R₇ ^(d), —C(O)OH, —C(O)OR₇^(e), —C(O)NR₁₀ ^(a)R₁₁ ^(a), —S(O)₂NR₁₀ ^(a)R₁₁ ^(a), or —S(O)_(p)R₇^(f); R₁ and R₂ are —H, an acyl group, a carbamoyl group, analkoxycarbonyl group, or an alkoxymethyl group; R₇ is —H, an optionallysubstituted alkyl, alkenyl, alkynyl, an optionally substitutedcycloalkyl, cycloalkenyl, an optionally substituted aryl, heteroaryl, anaralkyl, or an heteraralkyl; R₇ ^(a) is an optionally substituted alkyl,alkenyl, alkynyl, an optionally substituted cycloalkyl, cycloalkenyl,aryl, an aralkyl, or a heteraralkyl; R₇ ^(b) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, aryl, or heteroaryl; R₇ ^(b′) isalkyl, alkenyl, alkynyl, cycloalkyl, aralkyl, or heteraralkyl; R₇ ^(c)is an optionally substituted alkyl, alkenyl, alkynyl, an optionallysubstituted cycloalkyl, cycloalkenyl, an aralkyl, or a heteraralkyl; R₇^(d) is alkyl, aryl, or heteroaryl; R₇ ^(e) is alkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, an aralkyl, or a heteraralkyl; R₇ ^(f) is anoptionally substituted alkyl, alkenyl, alkynyl, an optionallysubstituted cycloalkyl, cycloalkenyl, an optionally substituted aryl,heteroaryl, an aralkyl, or a heteraralkyl; R₁₀ and R₁₁, for eachoccurrence, is independently —H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, aryl, or heteroaryl; or R₁₀ and R₁₁, taken together withthe nitrogen to which they are attached, form an optionally substitutedheterocyclyl; R₁₀ ^(a) and R₁₁ ^(a), for each occurrence, isindependently —H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,aryl, or heteroaryl; Y is —OH, —SH, an optionally substituted acyloxygroup, an optionally substituted alkoxycarbonyloxy group, or anoptionally substituted carbamoyloxy group; provided that when Y is —OHor —SH at least one of R₁ and R₂ is not hydrogen; and p, for eachoccurrence, is independently 1 or
 2. 13. The compound of claim 12,wherein the compound is not3-(2,4-Dihydroxy-5-ethyl-phenyl)-4-(3-trifluoromethyl-phenyl)-5-carbamoyloxy-[1,2,4]triazole;3-(2,4-Dihydroxy-5-ethyl-phenyl)-4-(1-methyl-indol-4-yl)-5-carbamoyloxy-[1,2,4]triazole;3-(2,4-Dihydroxy-5-methoxy-phenyl)-4-(8-methoxy-quinolin-5-yl)-5-carbamoyloxy-[1,2,4]triazole;3-(2-Hydroxy-4-ethoxycarbonyoxy-5-methoxy-phenyl)-4-(1-isopropyl-benzoimidazol-4-yl)-5-hydroxy-[1,2,4]triazole;3-(2-Hydroxy-4-ethoxycarbonyoxy-5-ethyl-phenyl)-4-(naphthalin-2-yl)-5-hydroxy-[1,2,4]triazole;3-[2-Hydroxy-4-(dimethyl-carbamoyoxy)-5-ethyl-phenyl]-4-(naphthalin-2-yl)-5-hydroxy-[1,2,4]triazole;3-[2-Hydroxy-4-(dimethyl-carbamoyoxy)-5-chloro-phenyl]-4-(quinolin-5-yl)-5-mercapto-[1,2,4]triazole;3-[2-Hydroxy-4-(dimethyl-carbamoyoxy)-5-ethyl-phenyl]-4-(2,3-difluoro-phenyl)-5-mercapto-[1,2,4]triazole;or3-[2-Hydroxy-4-isobutyryloxy-5-ethyl-phenyl]-4-(1-methyl-benzo-imidazol-4-yl)-5-hydroxy-[1,2,4]triazole.14. The compound of claim 12, wherein when Y is —OH or —SH, R₁ and R₂are selected from the group consisting of an acyl group, a carbamoylgroup, an alkoxycarbonyl group, and an alkoxymethyl group.
 15. Thecompound of claim 12, wherein R₁ and R₂ are the same.
 16. The compoundof claim 12, wherein X is —OR₇ or —SR₇ with R₇ being pyridyl.
 17. Thecompound of claim 12, wherein X is OC(O)R₇ ^(a) with R₇ ^(a) beingphenyl.
 18. The compound of claim 12, wherein X is C(O)R₇ ^(d) with R₇^(d) being methyl, ethyl, tert-butyl, phenyl, or pyridyl.
 19. Thecompound of claim 12, wherein X is S(O)_(p)R₇ ^(f) with R₇ ^(f) beingpyridyl.
 20. The compound of claim 12, wherein X is morpholino group,piperidinyl group, or 4-methylpiperazine-1-yl group.
 21. The compound ofclaim 12, wherein X is —OR₇ or —SR₇ with R₇ being alkyl substituted withan acyl group or an alkoxycarbonyl group.
 22. The compound of claim 12,wherein R₁₀ and R₁₁, taken together with the nitrogen to which they areattached, form an optionally substituted heterocyclyl selected from thegroup consisting of morpholino, piperidinyl, and4-methylpiperazine-1-yl.
 23. The compound of claim 12, wherein R is anoptionally substituted naphthyl group.
 24. The compound of claim 12,wherein R is represented by the following formula:

wherein: R₃, for each occurrence, is independently a substituentselected from the group consisting of alkyl, alkenyl, an alkynyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, halo, cyano, nitro,—NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, OC(O)R₇, —C(O)NH₂, —NHC(O)R₇, —SR₇,—S(O)_(p)R₇, —NHS(O)₂R₇, or —S(O)₂NH₂.
 25. The compound of claim 12,wherein R is an optionally substituted indolyl.
 26. The compound ofclaim 12, wherein R is represented by the following formula:

wherein: R₄ is H, a halo, alkyl, alkoxy, or alkyl sulfanyl; and R₅ is H,alkyl, or alkylcarbonyl.
 27. The compound of claim 12, wherein X is aC1-C6 alkyl, a C1-C6 haloalkyl, a C1-C6 alkoxy, a C1-C6 haloalkoxy, aC1-C6 alkyl sulfanyl or a C3-C6 cycloalkyl.
 28. A pharmaceuticalcomposition comprising: a pharmaceutically acceptable carrier and acompound according to claim 12.